Vesicular formulations

ABSTRACT

Disclosed herein are vesicular formulations that include one or more phospholipids and one or more surfactants and in certain embodiments the use of such formulations for the delivery of fatty acids for the treatment of disorders such as, fatty acid metabolic disorders, including essential fatty acid deficiency; pain or inflammation or osteoarthritis, more specifically for the treatment of deep tissue pain; asthma, bronchospasm, atherothrombatic cardiovascular disorders, avenous thrombatic disorders, inflammatory dermatoses disorders (e.g., atopic eczema, dishydrotic hand eczema, plaque type psoriasis, seborrheic eczema, and acne vulgaris), and dysmenorrhea.

This application claims the benefit of U.S. Provisional Application No.61/235,992, filed Aug. 21, 2009, U.S. Provisional Application No.61/314,476, filed Mar. 16, 2010, and U.S. Provisional Application No.61/320,154, filed Apr. 1, 2010, each of which are herein incorporated byreference in their entirety.

FIELD OF INVENTION

The present invention relates to formulations of phospholipids andsurfactants and to the use of such formulations for the delivery offatty acids for the treatment of disorders such as, fatty acid metabolicdisorders, including essential fatty acid deficiency; pain orinflammation or osteoarthritis, more specifically for the treatment ofdeep tissue pain; asthma, bronchospasm, atherothrombatic cardiovasculardisorders, avenous thrombatic disorders, inflammatory dermatosesdisorders (e.g., atopic eczema, dishydrotic hand eczema, plaque typepsoriasis, seborrheic eczema, and acne vulgaris), and dysmenorrhea.

BACKGROUND

U.S. Pat. No. 6,165,500 to Cevc describes a “preparation for theapplication of agents . . . provided with membrane-like structuresconsisting of one or several layers of amphiphilic molecules, or anamphiphilic carrier substance, in particular for transporting the agentinto and through natural barriers such as skin and similar materials.”Abstract. These transfersomes “consist of one or severalcomponents[,m]ost commonly a mixture of basic substances, one or severaledge-active substances, and agents [1.” Col. 5, lines 28-30. Accordingto U.S. Pat. No. 6,165,500, “l]ipids and other amphiphiles are bestsuited basic substances; surfactants or suitable solvents are the bestchoice from the point of view of edge-active substances[, and a]ll ofthese can be mixed with agents in certain proportions depending both onthe choice of the starting substances and on their absoluteconcentration.” Col. 5, lines 30-35.

U.S. Patent Application Publication No. US 2004/0071767 to Cevc et al.describes “formulations of nonsteroidal anti-inflammatory drugs (NSAIDs)based on complex aggregates with at least three amphiphatic componentssuspended in a . . . pharmaceutically acceptable . . . medium.”Abstract. “One of these components is capable of forming stable, largebilayer membranes on its own. The other at least two amphiphaticcomponents, including an NSAID. tend to destabilise such membranes,”Paragraph [0002].

U.S. Patent Application Publication No. US 2004/0105881 to Cevc et al.describes extended surface aggregates, “suspendable in a suitable liquidmedium and comprising at least three amphiphats (amphiphatic components)and being capable to improve the transport of actives throughsemi-permeable barriers, such as the skin, especially for thenon-invasive drug application in vivo by means of barrier penetration bysuch aggregates.” Paragraph [0002]. “The three amphiphats include atleast one membrane forming compound (MFC), which can form the membraneof [the aggregates], and at least two membrane destabilising compounds(MDC₁ and MDC₂) differentiated by their capability of forming smalleraggregates (with no extended surfaces) by either themselves or else incombination with each other and/or characterized by their relativelyhigh solubility in [the] suitable liquid medium. Paragraph [0002]. US2004/0105881 specifically discloses that “incorporation of a surfactantinto a bilayer membrane that is built from another less solubleamphiphat, such as a phospholipid, can increase the flexibility of theresulting complex membrane . . . promot[ing] the capability of complexaggregates . . . to cross pores in a semi-permeable membrane thatotherwise would prevent comparably large aggregates from crossing.”Paragraph [0015]. Citation of any reference in this section of theapplication is not an admission that the reference is prior art to theapplication. The above noted publications are hereby incorporated byreference in their entirety.

SUMMARY OF THE INVENTION

The invention encompasses vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effectivefor the delivery of fatty acids and/or phospholipids in the treatment ofdisorders related to fatty acid metabolic disorders, including essentialfatty acid deficiency. In a particular embodiment, the vesicularformulations comprise about 25% to about 30% surfactant by weight basedon the total weight of the vesicular formulation. These vesicularformulations are suitable for any method of administration, e.g ,subcutaneously, topically, or intravenously.

In accordance with the present invention, the vesicular formulations ofthe invention are formulated to deliver fatty acids and phosphatidylderivatives of fatty acids, such as arachidonic acid or omega-3 oromega-6 fatty acids. In accordance with this embodiment the vesicularformulations may optionally be formulated to include other lipidsdescribed herein, such as phosphatidyl choline, and surfactants. Inaccordance with the present invention, the vesicular formulations of theinvention deliver essential fatty acids, such as omega-3 fatty acids, todecrease the levels of triglycerides. Vesicular formulations of theinvention which deliver essential fatty acids, such as omega-3 fattyacids, may be useful in the treatment of fatty acid metabolic disorders,such as essential fatty acid deficiency and hypertriglyceridemia. In oneembodiment, the vesicular formulations which deliver essential fattyacids comprise a phosphatidylcholine derivative of a fatty acid, e.g., aphosphatidylcholine derivative of an omega-3 fatty acid. In variousembodiments, the vesicular formulation is one of the formulations setforth in Example Formulations 1-129.

“Fatty acid metabolic disorder” means a defect in one of the enzymesinvolved in fatty acid metabolism, and include fatty oxidationdisorders, whereby the body is unable to oxidize and metabolize fattyacids due to a failure in the enzymatic pathway. “Essential fatty aciddeficiency” means a deficiency in the essential fatty acids, e.g.,omega-3 and omega-6 fatty acids, which can lead to physical symptomssuch as hemorrhagic dermatitis skin atrophy, scaly dermatitis, dry skin,weakness, impaired vision, tingling sensations, mood swings, edema, highblood pressure, high triglycerides, hemorrhagic folliculitis,hematologic disturbances, immune and mental deficiencies, and impairedgrowth.

In accordance with the present invention, the vesicular formulations ofthe invention deliver fats and fat soluble vitamins, such as vitamin E,for the treatment of disorders related to hypolididemia, including,abetalipoproteinemia, hypobetalipoproteinemia, chlyomicron retentiondisease. In one embodiment, the vesicular formulations which deliverfats and fat soluble vitamins comprise a phosphatidylcholine derivativeof a fatty acid, e.g., a phosphatidylcholine derivative of vitamin E. Invarious embodiments, the vesicular formulation is one of theformulations set forth in Example Formulations 1-129.

The invention encompasses vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering organic matter once delivered to the subject. Thesevesicular formulations are suitable for any method of administration,e.g., subcutaneously, topically, or intravenously. Without in any waybeing limited by theory, it is believed that the surprisingly effectiveand capacious sequestration of native organic compounds by the vesicularformulations disclosed herein occurs because of liquid crystallinity ofthe vesicular formulations mediated by the presence of membrane adapterssuch as surfactants. In various embodiments, the vesicular formulationis one of the formulations set forth in Example Formulations 1-129. In apreferred embodiments, the vesicular formulations comprise about 25% toabout 30% surfactant by weight.

In one embodiment, the invention encompasses vesicular formulations oflipids and surfactants capable of sequestering native organic compounds,including arachidonic acid, upon delivery to human skin for thetreatment of pain or inflammation. In some embodiments, theseformulations are designed such that the vesicles are able to penetratedeep tissue without diversion into the blood vessels. That is, theformulations are able to travel to the site of the pain in sufficientamount to alleviate that pain to some extent. In accordance with theinvention, delivery to the deep tissue includes delivery of theformulation beneath the skin to the muscle tissue and to the jointitself, while limiting systemic delivery and exposure to theformulation. In a particular embodiment, the vesicular formulation iscapable of sequestering arachidonic acid upon administration to humanskin, and therefore is capable of altering the pathology of, e.g., painor inflammation. In another embodiment of the invention, vesicularformulations designed to sequester arachidonic acid may also be used toprevent the formation of metabolites, including eicosanoids, for theprevention and/or treatment of asthma, seborrheic eczema, bronchospasm,atherothrombatic cardiovascular disorders, venous thrombatic disorders,pain, and dysmenorrhea. In various embodiments, the vesicularformulation capable of sequestering arachidonic acid is one of theformulations set forth in Example Formulations 1-129.

In an embodiment of the invention, vesicular formulations comprising oneor more phospholipids and one or more nonionic surfactants can sequestercholesterol upon administration to human skin, thus decreasing theaccumulation of or uptake of cholesterol for the treatment ofhypercholesterolemia. The invention relates to vesicular formulationscomprising one or more phospholipids and one or more nonionicsurfactants that are effective in sequestering triglycerides, thusdecreasing the accumulation or uptake of triglycerides for the treatmentof hypertriglyceridemia. In various embodiments, the vesicularformulation capable of sequestering cholesterol or triglycerides is oneof the formulations set forth in Example Formulations 1-129.

The vesicular formulations of the invention may also be used tosequester factors involved in fatty acid metabolism, such as hormonesensitive lipase (HSL), Inhibition of HSL inhibits the conversion oftriglycerides to glycerol and fatty acids, resulting in a decrease inplasma free fatty acids. Thus, the vesicular formulations of theinvention have utility where the decrease in plasma fatty acids isdesired, including insulin resistance, metabolic syndrome X,dyslipidemias and abnormal lipoprotein metabolism. In variousembodiments, the vesicular formulation capable of sequestering factorsinvolved in fatty acid metabolism is one of the formulations set forthin Example Formulations 1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that arc effective insequestering metal (as a chelator) upon administration to human skin forthe treatment of, e.g., metal toxicity. In various embodiments, thevesicular formulation capable of sequestering metals is one of theformulations set forth in Example Formulations 1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering a toxin (e.g., DDT) upon administration to human skin. Invarious embodiments, the vesicular formulation capable of sequesteringtoxins such as DDT is one of the formulations set forth in ExampleFormulations 1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering inflammatory mediators (e.g., cytokines, such asinterleukins, or presenting antigens) upon administration to human skinfor the treatment of inflammation and inflammatory related disorders,such as asthma. In various embodiments, the vesicular formulationcapable of sequestering inflammatory mediators is one of theformulations set forth in Example Formulations 1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering amyloid upon administration for the treatment ofAlzheimer's disease. Such formulations may be administered intravenouslyin accordance with this embodiment. In various embodiments, thevesicular formulation capable of sequestering amyloid is one of theformulations set forth in Example Formulations 1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering uric acid upon administration for the treatment of gout ormacular degeneration, such as AMD. Such formulations may be administeredtopically or intravitreally in accordance with this embodiment. Invarious embodiments, the vesicular formulation capable of sequesteringuric acid is one of the formulations set forth in Example Formulations1-129.

The invention relates to vesicular formulations comprising one or morephospholipids and one or more nonionic surfactants that are effective insequestering squalene, thus leading to fungistatic activity against,e.g., hyphal fungi. These vesicular formulations are suitable for anymethod of administration, e.g., subcutaneously, topically, orintravenously. In various embodiments, the vesicular formulation capableof sequestering squalene is one of the formulations set forth in ExampleFormulations 1-129.

The formulations of the invention are formulated in the absence of anypharmaceutically active agent, i.e., any non-lipid non-surfactantpharmaceutically active agent.

As used herein, the term “formulation” is not meant to imply that theingredients or components are in combination with a pharmaceuticallyactive agent, i.e., any non-lipid non-surfactant active agent that hasreceived regulatory approval for the treatment of fatty acid relateddisorders, hypocholesterolemia, hypertriclyceridemia, pain, includingosteoarthritic pain, inflammation, infection, or toxicity, includingmetal toxicity or any of the disorders listed above.

Despite the lack of a recognized active agent, the vesicles elicit atherapeutic effect, namely the treatment of disorders related to fattyacid deficiencies, fatty acid metabolism, hypertriglyceridemia andhypercholesterolemia, or any of the disorders listed above. Withoutbeing bound by any theory, Applicant believes that the vesiclecomponents themselves are responsible for this effect.

In one embodiment, the invention provides a pharmaceutical package orkit comprising one or more containers filled with the formulation of theinvention, and instructions for administration of the formulation to apatient or subject in need thereof for the treatment of any disordersrelated to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia and hypercholesterolemia, or any of the disorderslisted above. In certain embodiments, the formulation comprises one ormore phospholipids and one or more surfactants. In certain embodiments,the formulation does not comprise a pharmaceutically active agent, i.e.,any non-lipid, non-surfactant pharmaceutically active agent that hasreceived marketing or regulatory approval in any country for thetreatment of disorders related to fatty acid deficiencies, fatty acidmetabolism, hypertriglyceridemia, hypercholesterolemia, pain, includingosteoarthritic pain, inflammation, infection, including fungal orbacterial infection, or toxicity, including metal toxicity, or any ofthe other disorders listed above. In various embodiments, the containercomprises a formulation formulated as a suspension, emulsion, gel,cream, lotion, spray, film forming solution or lacquer. The inventionprovides packages or kits that can be used in any of the above-describedmethods.

In one embodiment, the invention comprises a method for the treatment ofdisorders related to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia, hypercholesterolemia, or any of the disorderslisted above, wherein the vesicular formulations of the invention areadministered over a period of one or more weeks, for example for atleast five weeks, six weeks, seven weeks, eight weeks, nine weeks, tenweeks, eleven weeks, or twelve weeks, sixteen weeks, twenty four weeks,four months, six months, eight months, ten months, one year, two or moreyears, or indefinitely.

In one embodiment, the formulations of the invention comprise one ormore phospholipids, one or more nonionic surfactants, in the absence ofany pharmaceutically active agent, i.e., any non-lipid non-surfactantpharmaceutically active agent that has received regulatory approval forthe treatment of disorders related to fatty acid deficiencies, fattyacid metabolism, hypertriglyceridemia, hypercholesterolemia pain,including osteoarthritic pain, inflammation, infection, including fungalor bacterial infection, or toxicity, including metal toxicity, or any ofthe disorders listed above.

In one embodiment, a 0.1 to 10 gram dose of the formulation of theinvention is administered to the patient for the treatment of disordersrelated to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia and hypercholesterolemia, or any of the disorderslisted above; a 1 to 10 gram dose of the formulation is administered tothe patient for the treatment of disorders related to fatty aciddeficiencies, fatty acid metabolism, hypertriglyceridemia,hypercholesterolemia, or any of the disorders listed above; a 1 to 5gram dose of the formulation is administered to the patient for thetreatment of disorders related to fatty acid deficiencies, fatty acidmetabolism, hypertriglyceridemia, hypercholesterolemia, pain, includingosteoarthritic pain, inflammation, infection, including fungal orbacterial infection, or toxicity, including metal toxicity, or any ofthe disorders listed above; or a 1 gram, 2 gram, 3 gram, 4 gram, 5 gram,6 gram, 7 gram, 8 gram, 9 gram or 10 gram dose of the formulation isadministered to the patient for the treatment of disorders related tofatty acid deficiencies, fatty acid metabolism, hypertriglyceridemia,hypercholesterolemia pain, including osteoarthritic pain, inflammation,infection, including fungal or bacterial infection, or toxicity,including metal toxicity, or any of the disorders listed above. In someembodiments, the dose is measured as the total weight of thedeformasome. In some embodiments, the dose is measured as the totalweight of the lipid(s) and surfactant(s) in the deformasome. The dosemay be administered once or twice daily for the treatment of disordersrelated to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia and hypercholesterolemia, or any of the disorderslisted above. The dose may be administered once, twice, three, four,five, six, or seven times per week in accordance with the invention. Thedose may be administered every day, every other day, or two to threetimes a week in accordance with the invention.

In some embodiments, the lipid in the pharmaceutical composition is aphospholipid. In some embodiments, the second lipid is alysophospholipid. In some embodiments, the surfactant is a non-ionicsurfactant.

In some embodiments, the compositions of the invention form vesicles orother extended surface aggregates (ESAs), wherein the vesicularpreparations have improved permeation capability through thesemi-permeable barriers, such as skin. The adaptability anddeformability of the vesicles allow the vesicles to penetrate beneaththe skin to the muscle and the joint itself, however, the size of thevesicle prevents penetration into the vasculature and as a resultprevents systemic delivery. While not to be limited to any mechanism ofaction, the formulations of the invention are able to form vesiclescharacterized by their deformability and/or adaptability. Theadaptability or deformability of the vesicles may be determined by theability of the vesicles to penetrate a barrier with pores having anaverage pore diameter at least 50% smaller than the average vesiclediameter before the penetration.

In some embodiments, the vesicular compositions of the invention providefor targeted delivery of e.g., fatty acids to phospholipase-rich sites,e.g., tissues that are part of an inflammatory process or sitescontaining microorganisms such as bacteria (including narcadia) orfungi. While not to be limited to any mechanism of action or by anytheory, the vesicular compositions of the invention are broken down byphospholipases. Thus, phospholipases that are released as part of theinflammatory process (e.g., cancer or asthma) or that are released uponcontact with a microorganism such as bacteria or fungi can lead to anumber of effects including but not limited to rapid entry of thevesicular compositions into the target tissue, changes in theintracellular or intramembraneous lipid homeostasis, which may lead toincreased apoptosis or altered membrane function, including increasedpermeability, and rapid metabolism of the vesicular composition withrelease of its constituents.

DETAILED DESCRIPTION OF THE INVENTION

Generally, the nomenclature used herein and the laboratory procedures inorganic chemistry, medicinal chemistry, and pharmacology describedherein are those well known and commonly employed in the art. Unlessdefined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this disclosure belongs.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human), cow, sheep, goat, pig, horse, dog, cat, rabbit,rat, or mouse. The terms “subject” and “patient” are usedinterchangeably herein in reference, for example, to a mammaliansubject, such as a human subject.

As used herein, a “sufficient amount.” “amount effective to” or an“amount sufficient to” achieve a particular result refers to an amountof the formulation of the invention is effective to produce a desiredeffect, which is optionally a therapeutic effect (i.e., byadministration of a therapeutically effective amount). Alternativelystated, a “therapeutically effective” amount is an amount that providessome alleviation, mitigation, and/or decrease in at least one clinicalsymptom. Clinical symptoms associated with the disorder that can betreated by the methods of the invention are well-known to those skilledin the art. Further, those skilled in the art will appreciate that thetherapeutic effects need not be complete or curative, as long as somebenefit is provided to the subject. For example, a “sufficient amount”or “an amount sufficient to” can be an amount that is effective to treatthe symptoms of fatty acid deficiencies, hypertriglyceridemia orhypercholesterolemia or other joint or muscle pain.

As used herein, the terms “treat”, “treating” or “treatment of” meanthat the severity of a subject's condition is reduced or at leastpartially improved or ameliorated and/or that some alleviation,mitigation or decrease in at least one clinical symptom is achievedand/or there is an inhibition or delay in the progression of thecondition and/or delay in the progression of the onset of disease orillness. The terms “treat”, “treating” or “treatment of” also meansmanaging the disease state.

As used herein, the term “pharmaceutically acceptable” when used inreference to the formulations of the invention denotes that aformulation does not result in an unacceptable level of irritation inthe subject to whom the formulation is administered. Preferably suchlevel will be sufficiently low to provide a formulation suitable forapproval by regulatory authorities.

As used herein with respect to numerical values, the term “about” meansa range surrounding a particular numeral value which includes that whichwould be expected to result from normal experimental error in making ameasurement. For example, in certain embodiments, the term “about” whenused in connection with a particular numerical value means±20%, unlessspecifically stated to be ±1%, ±2%, ±3%, ±4%, ±5%, ±10%. ±15%, or ±20%of the numerical value.

The term “alkyl” refers to a linear or branched saturated monovalenthydrocarbon radical, wherein the alkyl may optionally be substitutedwith one or more substituents Q as described herein. The term “alkyl”also encompasses both linear and branched alkyl, unless otherwisespecified. In certain embodiments, the alkyl is a linear saturatedmonovalent hydrocarbon radical that has 1 20 (C₁₋₂₀), 1 to 15 (C₁₋₁₅), 1to 12 (C₁₋₁₂), 1 to 10 (C₁₋₁₀), or 1 to 6 (C₁₋₆) carbon atoms, or abranched saturated monovalent hydrocarbon radical of 3 to 20 (C₃₋₂₀), 3to 15 (C₃₋₁₅), 3 to 12 (C₃₋₁₂), 3 to 10 (C₃₋₁₀), or 3 to 6 (C₃₋₆) carbonatoms. As used herein, linear C₁₋₆ and branched C₃₋₆ alkyl groups arealso referred as “lower alkyl.” Examples of alkyl groups include, butare not limited to, methyl, ethyl, propyl (including all isomericforms), n-propyl, isopropyl, butyl (including all isomeric forms),n-butyl, isobutyl, sec-butyl, t-butyl, pentyl (including all isomericforms), and hexyl (including all isomeric forms). For example, C₁₋₆alkyl refers to a linear saturated monovalent hydrocarbon radical of 1to 6 carbon atoms or a branched saturated monovalent hydrocarbon radicalof 3 to 6 carbon atoms. It is understood in the chemical arts, that theuse of the longer chains described herein may be appropriate, orappropriate only in limited amounts, within a molecule so that theproperties of the resulting molecule (such as solubility) areappropriate for the use. Thus, while those in the art may use the abovelonger length alkyl substituents they will be used only when appropriateto provide the desired function.

The term “aryl” refers to a monocyclic aromatic group and/or multicyclicmonovalent aromatic group that contain at least one aromatic hydrocarbonring. In certain embodiments, the aryl has from 6 to 20 (C₆₋₂₀), from 6to 15 (C₆₋₁₅), or from 6 to 10 (C₆₋₁₀) ring atoms. Examples of arylgroups include, but are not limited to, phenyl, naphthyl, fluorenyl,azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Arylalso refers to bicyclic or tricyclic carbon rings, where one of therings is aromatic and the others of which may be saturated, partiallyunsaturated, or aromatic, for example, dihydronaphthyl, indenyl,indanyl, or tetrahydronaphthyl (tetralinyl). In certain embodiments,aryl may also be optionally substituted with one or more substituents Qas described herein.

The term “heteroaryl” refers to a monocyclic aromatic group and/ormulticyclic aromatic group that contain at least one aromatic ring,wherein at least one aromatic ring contains one or more heteroatomsindependently selected from O, S, and N. Each ring of a heteroaryl groupcan contain one or two O atoms, one or two S atoms, and/or one to four Natoms, provided that the total number of heteroatoms in each ring isfour or less and each ring contains at least one carbon atom. Theheteroaryl may be attached to the main structure at any heteroatom orcarbon atom which results in the creation of a stable compound. Incertain embodiments, the heteroaryl has from 5 to 20, from 5 to 15, orfrom 5 to 10 ring atoms. Examples of monocyclic heteroaryl groupsinclude, but are not limited to. pyrrolyl, pyrazolyl, pyrazolinyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl,furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl,pyridazinyl, and triazinyl. Examples of bicyclic heteroaryl groupsinclude, but are not limited to, indolyl, benzothiazolyl, benzoxazolyl,benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl,benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl,isobenzofuranyl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl,indazolyl, purinyl, pyrrolopyridinyl, furopyridinyl, thicnopyridinyl,dihydroisoindolyl, and tetrahydroquinolinyl. Examples of tricyclicheteroaryl groups include, but are not limited to, carbazolyl,benzindolyl, phenanthrollinyl, acridinyl, phenanthridinyl, andxanthenyl. In certain embodiments, heteroaryl may also be optionallysubstituted with one or more substituents Z as described herein.

The term “alkenoyl” as used herein refers to —C(O)-alkenyl. The term“alkenyl” refers to a linear or branched monovalent hydrocarbon radical,which contains one or more, in one embodiment, one to five,carbon-carbon double bonds. The alkenyl may be optionally substitutedwith one or more substituents Z as described herein. The term “alkenyl”also embraces radicals having “cis” and “trans” configurations, oralternatively, “Z” and “E” configurations, as appreciated by those ofordinary skill in the art. As used herein, the term “alkenyl”encompasses both linear and branched alkenyl, unless otherwisespecified. For example, C₂₋₆ alkenyl refers to a linear unsaturatedmonovalent hydrocarbon radical of 2 to 6 carbon atoms or a branchedunsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms. Incertain embodiments, the alkenyl is a linear monovalent hydrocarbonradical of 2 to 30 (C₂₋₃₀), 2 to 20 (C₂₋₂₀), 2 to 20 (C₂₋₂₀), 2 to 15(C₂₋₁₅), 2 to 12 (C₂₋ ₁₂), 2 to 10 (C₂₋₁₀, or 2 to 6 (C₂₋₆) carbonatoms, or a branched monovalent hydrocarbon radical of 3 to 30 (C₃₋₃₀),3 to 24 (C₃₋₂₄), 3 to 20 (C₃₋₂₀), 3 to 15 (C₃₋₁₅), 3 to 12 (C₃₋₁₂), 3 to10 (C₃₋₁₀), or 3 to 6 (C₃₋₆) carbon atoms. Examples of alkenyl groupsinclude, but are not limited to, ethenyl, propen-1-yl, propen-2-yl,allyl, butenyl, and 4-methylbutenyl. In certain embodiments, thealkenoyl is mono-alkenoyl, which contains one carbon-carbon double bond.In certain embodiments, the alkenoyl is di-alkenoyl, which contains twocarbon-carbon double bonds. In certain embodiments, the alkenoyl ispoly-alkenoyl, which contains more than two carbon-carbon double bonds.

The term “heterocyclyl” or “heterocyclic” refers to a monocyclicnon-aromatic ring system and/or multicyclic ring system that contains atleast one non-aromatic ring, wherein one or more of the non-aromaticring atoms are heteroatoms independently selected from O, S, or N: andthe remaining ring atoms are carbon atoms. In certain embodiments, theheterocyclyl or heterocyclic group has from 3 to 20, from 3 to 15, from3 to 10, from 3 to 8, from 4 to 7, or from 5 to 6 ring atoms. In certainembodiments, the heterocyclyl is a monocyclic, bicyclic, tricyclic, ortetracyclic ring system, which may include a fused or bridged ringsystem, and in which the nitrogen or sulfur atoms may be optionallyoxidized, the nitrogen atoms may be optionally quaternized, and somerings may be partially or fully saturated, or aromatic. The heterocyclylmay be attached to the main structure at any heteroatom or carbon atomwhich results in the creation of a stable compound. Examples of suchheterocyclic radicals include, but are not limited to, acridinyl,azepinyl, benzimidazolyl, benzindolyl, benzoisoxazolyl, benzisoxazinyl,benzodioxanyl, benzodioxolyl, benzofuranonyl, benzofuranyl,benzonaphthofuranyl, benzopyranonyl, benzopyranyl,benzotetrahydrofuranyl, benzotetrahydrothienyl, benzothiadiazolyl,benzothiazolyl, benzothiophenyl, benzotriazolyl, benzothiopyranyl,benzoxazinyl, benzoxazolyl, benzothiazolyl, β-carbolinyl, carbazolyl,chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl,dibenzofuranyl, dihydrobenzisothiazinyl, dihydrobenzisoxazinyl,dihydrofuryl, dihydropyranyl, dioxolanyl, dihydropyrazinyl,dihydropyridinyl, dihydropyrazolyl, dihydropyrimidinyl, dihydropyrrolyl,dioxolanyl, 1,4-dithianyl, furanonyl, furanyl, imidazolidinyl,imidazolinyl, imidazolyl, imidazopyridinyl, imidazothiazolyl, indazolyl,indolinyl, indolizinyl, indolyl, isobenzotetrahydrofuranyl,isobenzotetrahydrothienyl, isobenzothienyl, isochromanyl, isocoumarinyl,isoindolinyl, isoindolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl,isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl,octahydroindolyl, octahydroisoindolyl, oxadiazolyl, oxazolidinonyl,oxazolidinyl, oxazolopyridinyl, oxazolyl, oxiranyl, perimidinyl,phenanthridinyl, phenathrolinyl, phenarsazinyl, phenazinyl,phenothiazinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl,4-piperidonyl, pteridinyl, purinyl, pyrazinyl, pyrazolidinyl, pyrazolyl,pyridazinyl, pyridinyl, pyridopyridinyl, pyrimidinyl, pyrrolidinyl,pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl,quinuclidinyl, tetrahydrofuryl, tetrahydrofuranyl,tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl,tetrazolyl, thiadiazolopyrimidinyl, thiadiazolyl, thiamorpholinyl,thiazolidinyl, thiazolyl, thienyl, triazinyl, triazolyl, and1,3,5-trithianyl. In certain embodiments, heterocyclic may also beoptionally substituted with one or more substituents Z as describedherein.

The term “halogen”, “halide” or “halo” refers to fluorine, chlorine,bromine, and/or iodine.

The term “optionally substituted” is intended to mean that a group,including alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl,heteroaryl, and heterocyclyl, may be substituted with one or moresubstituents Z, in one embodiment, one, two, three or four substituentsZ, where each Z is independently selected from the group consisting ofcyano, halo, oxo, nitro, C₁₋₆ alkyl, halo-C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, C₃₋₇ cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₄ aralkyl, heteroaryl,heterocyclyl, —C(O)R^(e), —C(O)OR^(e), —C(O)NR^(f)R_(g),—C(NR^(e))NR^(f)R_(g), —OR^(e), —OC(O)R^(e), —OC(O)OR^(e),—OC(O)NR^(f)R^(g), —OC(═NR^(e))NR^(f)R^(g), —OS(O)R^(e), —OS(O)₂R^(e),—OS(O)NR^(f)R^(g), —OS(O)₂NR^(f)R^(g), —NR^(f)R^(g), —NR^(e)C(O)R^(f),—NR^(e)C(O)OR^(f), —NR^(e)C(O)NR^(f)R^(g), —NR^(e)C(═NR^(h))NR^(f)R^(g),—NR^(e)S(O)R^(f), —NR^(e)S(O)₂R^(f), —NR^(e)S(O)NR^(f)R^(g),—NR^(e)S(O)₂NR^(f)R^(g), —SR^(e), —S(O)R^(e), —S(O)₂R^(e), and—S(O)₂NR^(f)R^(g), wherein each R^(e), R^(f), R^(g), and R^(h) isindependently hydrogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇cycloalkyl, C₆₋₁₄ aryl, C₇₋₁₄ aralkyl, heteroaryl, or heterocyclyl; orR^(f) and R^(g) together with the N atom to which they are attached formheterocyclyl.

The term “solvate” refers to a compound provided herein or a saltthereof, which further includes a stoichiometric or non-stoichiometricamount of solvent bound by non-covalent intermolecular forces. Where thesolvent is water, the solvate is a hydrate.

In accordance with this disclosure, the term “comprising” is inclusiveor open-ended and does not exclude additional, unrecited elements ormethod steps; the term “consisting of” excludes any element, step, oringredient not specified; and the term “consisting essentially of”excludes any element, step, or ingredient that materially changes abasic characteristic of the invention.

In some embodiments, the formulation of the invention provided hereincomprise at least one lipid, preferably a phospholipid, at least onesurfactant, preferably a nonionic surfactant, optionally suspended in apharmaceutically acceptable medium, preferably an aqueous solution,preferably having a pH ranging from 3.5 to 9.0, preferably from 4 to7.5. The formulation of the invention may optionally contain buffers,antioxidants, preservatives, microbicides, antimicrobials, emollients,co-solvents, and/or thickeners. In some embodiments, the formulation ofthe invention comprises a mixture of more than one lipid, preferablymore than one phospholipids. In some embodiments, the formulation of theinvention consists essentially of at least one lipid, preferably aphospholipid, at least one surfactant, preferably a nonionic surfactant,a pharmaceutically acceptable carrier, and optionally buffers,antioxidants, preservatives, microbicides, antimicrobials, emollients,co-solvents, and/or thickeners. In some embodiments, the formulation ofthe invention consists of at least one lipid, preferably a phospholipid,at least one surfactant, preferably a nonionic surfactant, apharmaceutically acceptable carrier, and one or more of the following:buffers, antioxidants, preservatives, microbicides, antimicrobials,emollients, co-solvents, and thickeners.

LIPID

In the sense of this disclosure, a “lipid” is any substance, which hasproperties like or similar to those of a fat. As a rule, it has anextended apolar group (the “chain”, X) and generally also awater-soluble, polar hydrophilic part, the “head” group (Y) and has thebasic Formula I:

X-Y_(n)  (I)

wherein n is equal to or larger than zero.

Lipids with n=0 are referred to as apolar lipids and lipids with n≧1 arereferred to as polar lipids. In this sense, all amphiphilic substances,including, but not limited to glycerides, glycerophospholipids,glycerophosphinolipids, glycerophosphonolipids, sulfolipids,sphingolipids, isoprenoid lipids, steroids or sterols andcarbohydrate-containing lipids can generally be referred to as lipids,and are included as such in this disclosure. A list of relevant lipidsand lipid related definitions is provided in EP 0 475 160 A1 (see, e.g.p. 4,1. 8 to p. 6,1. 3) and U.S. Pat. No. 6,165,500 (see, e.g., col.6,1. 10 to col. 7,1. 58), each incorporated herein by reference in theirentirety.

A phospholipid in various embodiments may contain (1) a moiety derivedfrom glycerol or a sphingosine, (2) a phosphate group, and/or (3) simpleorganic molecule such as choline. A phospholipid as used herein may, forexample. be a compound of Formula II:

R¹—CH₂—CHR²—CR³H—O—PHO₂—O—R⁴  (II)

wherein R¹ and R² are hydrogen, OH, an alkyl group, an aliphatic chain,an aliphatic chain derived from a fatty acid or a fatty alcohol:provided however that R¹ and R² cannot both be hydrogen, OH or a C₁-C₃alkyl group, In some embodiments R¹ and R² are independently, analiphatic chain, most often derived from a fatty acid or a fattyalcohol; R³ generally is a hydrogen.

The OH-group of the phosphate is a hydroxyl radical or hydroxyl anion(i.e., hydroxide) form, dependent on degree of the group ionization.Furthermore, R⁴ may he a proton or a short-chain alkyl group,substituted by a tri-short-chain alkylammonium group. such as atrimethylammonium group, or an amino-substituted short-chain alkylgroup, such as 2-trimethylammonium ethyl group (cholinyl) or2-dimethylammonium short alkyl group.

A sphingophospholipid is, for example, a compound of Formula IIB:

R¹-Sphingosine-O—PHO₂—O—R⁴  (IIB)

wherein R¹ is a fatty-acid attached via an amide bond to the nitrogen ofthe sphingosine and R⁴ has the meanings given under Formula II.

A lipid preferably is a substance of formulae II or IIB, wherein R¹and/or R² are acyl or alkyl, n-hydroxyacyl or n-hydroxyalkyl, but mayalso be branched, with one or more methyl groups attached at almost anypoint of the chain; usually, the methyl group is near the end of thechain (iso or anteiso). The radicals R¹ and R² may moreover either besaturated or unsaturated (mono-, di- or poly-unsaturated). R³ ishydrogen and R⁴ is 2-trimethylammonium ethyl (the latter corresponds tothe phosphatidyl choline head group), 2-dimethylammonium ethyl,2-methylammonium ethyl or 2-aminoethyl (corresponding to thephosphatidyl ethanolamine head group). R⁴ may also be a proton (givingphosphatidic acid), a serine (giving phosphatidylserine), a glycerol(giving phosphatidylglycerol), an inositol (givingphosphatidylinositol), or an alkylamine group (givingphosphatidylethanolamine in case of an ethylamine), if one chooses touse a naturally occurring glycerophospholipid. Otherwise, any othersufficiently polar phosphate ester, such that will form a lipid bilayer,may be considered as well for making the formulations of the disclosure.

A phospholipid is, for example, a compound of Formula IIC:

wherein R¹ and R² are independently an acyl group, alkyl group,n-hydroxyacyl group, or n-hydroxyalkyl group, most often derived from afatty acid or a fatty alcohol, wherein R¹ and R² may also be branched,with one or more methyl groups attached at almost any point of thechain: usually, the methyl group is near the end of the chain (iso oranteiso), wherein R¹ and R² cannot both be hydrogen, OH or a C₁-C₃ alkylgroup. The radicals R¹ and R² may moreover either be saturated orunsaturated (mono-, di- or poly-unsaturated). R³ generally is ahydrogen. The OH-group of the phosphate is a hydroxyl radical orhydroxyl anion (i.e., hydroxide) form, dependent on degree of the groupionization. Furthermore, R⁴ may be a proton or a short-chain alkylgroup, substituted by a tri-short-chain alkylammonium group, such as atrimethylammonium group, or an amino-substituted short-chain alkylgroup, such as 2-trimethylammonium ethyl group (cholinyl) or2-dimethylammonium short alkyl group. R⁴ may be 2-trimethylammoniumethyl (the latter corresponds to the phosphatidyl choline head group),2-dimethylammonium ethyl, 2-methylammonium ethyl or 2-aminoethyl(corresponding to the phosphatidyl ethanolamine head group). R⁴ may alsobe a proton (giving phosphatidic acid), a serine (givingphosphatidylserine), a glycerol (giving phosphatidylglycerol), aninositol (giving phosphatidylinositol), or an alkylamine group (givingphosphatidylethanolamine in case of an ethylamine), if one chooses touse a naturally occurring glycerophospholipid. Otherwise, any othersufficiently polar phosphate ester, such that will form a lipid bilayer,may be considered as well for making the formulations of the disclosure.

Table 1 lists preferred phospholipids in accordance with one embodimentof the disclosure.

TABLE 1 Preferred (phospho)lipids Phospholipid: Type and Charge Fattychain Phosphatidyl- Phosphatidyl- Phosphatidyl- Phosphatidyl-Phosphatidic Length: nr. of choline/± ethanolamine/± Sphingomyelin/+glycerol/− inositol/− acid/− Name(s) double bonds Main lipid, L1 Mainlipid, L1 Main lipid, L1 Aux. lipid, L2 Aux. lipid, L2 Aux. lipid, L2C24 Behen(o)yl C22 Eruca(o)yl C22:1-13cis Arachin(o)yl C20 Gadolen(o)ylC20:1-11cis Arachidon(o)yl C20:4-5,8,11,14cis Ole(o)yl C18:1-9cis DOPCDOPE SM-oleyl DOPG DOPI DOPA Stear(o)yl C18 Linol(o)yl C18:2-9,12cis(Soy-PC/ (Soy-PE/ Brain SM (Soy-PC/ (Soy-PI/ (Soy-PA/ Linole(n/o)ylC18:3-9,12,15cis Egg-PC) Egg-PE) Egg-PC) Liver-PI) Egg-PA)Palmitole(o)yl C18:1-9cis Palmit(o)yl C16 Myrist(o)yl C14 DMPC DMPESM-myristyl DMPG DMPI Laur(o)yl C12 DLPC DLPE SM-lauryl DLPA Capr(o)ylC10 Rel concentration range L1/L2 (M/M) 1/0 1/0 10/1-1/1 10/1-3/110/1-5/1 “Total Lipid”* concentration range (w-%) 0.5-45 0.5-45  0.5-40 0.5-40  0.5-40 *Total Lipid includes phospholipid(s), surfactant(s)tand all lipophilic excipients

The preferred lipids in the context of this disclosure are uncharged andform stable, well hydrated bilayers; phosphatidylcholines,phosphatidylethanolamine, and sphingomyelins are the most prominentrepresentatives of such lipids. Any of those can have chains as listedin the Table 1, the ones forming fluid phase bilayers, in which lipidchains are in disordered state, being preferred.

Different negatively charged, i.e., anionic, lipids can also beincorporated into vesicular lipid bilayers. Attractive examples of suchcharged lipids are phosphatidylglycerols, phosphatidylinositols and,somewhat less preferred, phosphatidic acid (and its alkyl ester) orphosphatidylserine. It will be realized by anyone skilled in the artthat it is less commendable to make vesicles just from the chargedlipids than to use them in a combination with electro-neutral bilayercomponent(s). In case of using charged lipids, buffer composition and/orpH care must selected so as to ensure the desired degree of lipidhead-group ionization and/or the desired degree of electrostaticinteraction between the, oppositely, charged drug and lipid molecules.Moreover, as with neutral lipids, the charged bilayer lipid componentscan in principle have any of the chains listed in the Table 1. Thechains forming fluid phase lipid bilayers are clearly preferred,however, both due to vesicle adaptability increasing role of increasingfatty chain fluidity and due to better ability of lipids in fluid phaseto mix with each other.

The fatty acid- or fatty alcohol-derived chain of a lipid is typicallyselected amongst the basic aliphatic chain types given in the followingtables:

TABLE 2 The (most) preferred basic, straight, saturated fatty chainresidues Shorthand designation Systematic name Trivial name 12:0Dodecanoic Lauric 13:0 Tridecanoic 14:0 Tetradecanoic Myristic 15:0Pentadecanoic 16:0 Hexadecanoic Palmitic 17:0 Heptadecanoic Margaric18:0 Octadecanoic Stearic 19:0 Nonadecanoic 20:0 Eicosanoic Arachidic21:0 Heneicosanoic 22:0 Docosanoic Behenic 23:0 Tricosanoic 24:0Tetracosanoic Lignoceric

TABLE 3 The (most) preferred monoenoic fatty chain residues Shorthanddesignation Systematic name Trivial name 9-14:1/14:1(n − 5)cis-9-Tetradecenoic Myristoleic 7-16:1/16:1(n − 9) cis-7-Hexadecenoic9-16:1/16:1(n − 7) cis-9-Hexadecenoic Palmitoleic 9-18:1/18:1(n − 9)cis-9-Octadecenoic Oleic 11-18:1/18:1(n − 7) cis-11-Octadecenoiccis-Vaccenic 11-20:1/20:1(n − 9) cis-11-Eicosenoic Gondoic14-20:1/20:1(n − 6) cis-14-Eicosaenoic 13-22:1/22:1(n − 9)cis-13-Docosenoic Erucic 15-24:1/24:1(n − 9) cis-15-TetracosenoicNervoni 3t-18:1 trans-3-Hexadecenoi 9t-18:1 trans-9-Octadecenoic Elaidic11t-18:1 trans-11-Octadecenoic Vaccenic

TABLE 4 The (most) preferred dienoic and polyenoic fatty chain residuesShorthand designation Systematic name Trivial name 10,13c-16:2/16:2(n −3) 10-cis,13-cis-Hexadecadienoic 7,10c-16:2/16:3(n − 6)7-cis,10-cis-Hexadecadienoic 7,10,13c-16:3/16:3(n − 3)7-cis,10-cis,13-cis-Hexadecatrienoic 12,15c-18:2/18:2(n − 3)12-cis,15-cis-Octadecadienoic α-Linoleic 10,12t-18:2/18:2(n − 6)trans-10,trans-12-Octadecadienoic 9,12c-18:2/18:2(n − 6)9-cis,12-cis-Octadecadienoic γ-Linoleic 9,12,15c-18:3/18:3(n − 3)9-cis,12-cis,15-cis-Octadecatrienoic α-Linolenic 6,9,12c-18:3/18:3(n −6) 6-cis,9-cis,12-cis-Octadecatrienoic γ-Linolenic 9c,11c,13t-18:39-cis,11-trans,13-trans-Octadecatrienoic α-Eleostearic 8t,10t,12c-18:38-trans,10-trans,12-cis-Octadecatrienoic Calendic 6,9,12,15c-18:4/18:4(n− 3) 6,9,12,15-Octadecatetraenoic Stearidonic 3,6,9,12c-18:4/18:4(n − 6)3,6,9,12-Octadecatetraenoic 3,6,9,12,15c-18:5/18:5(n − 3)3,6,9,12,15-Octadecapentaenoic 14,17c-20:2/20:2(n − 3)14-cis,17-cis-Eicosadienoic 11,14c-20:2/20:2(n − 6)11-cis,14-cis-Eicosadienoic 11,14,17c-20:3/20:3(n − 3)8-cis,11-cis,14-cis-Eicosatrienoic Dihomo-α-linolenic8,11,14c-20:3/20:3(n − 6) 8-cis,11-cis,14-cis-EicosatrienoicDihomo-γ-linolenic 5,8,11c-20:3 20:3(n − 9) 5,8,11all-cis-Eicosatrienoic‘Mead's’ 5,8,11,14c-20:4/20:4(n − 6) 5,8,11;14-all-cis-EicosatetraenoicArachidonic 8,11,14,17c-20:4/20:4(n − 3)8,11,14,17-all-cis-Eicosatetraenoic 5,8,11,14,17c-20:5 or 20:5(n − 3)5,8,11,14,17-all-cis-Eicosapentaenoic 13,16c-22:2 13,16-Docosadienoic13,16,19c-22:3/22:3(n − 3) 13,16,19-Docosatrienoic 10,13,16c-22:3/22:3(n− 6) 10,13,16-Docosatrienoic 7,10,13,16c-22:4/22:4(n − 6)7,10,13,16-Docosatetraenoic Adrenic 4,7,10,13,16c-22:5 or 22:5(n − 6)4,7,10,13,16-Docosapentaenoic 4,7,10,13,16,19c-22:5 or 22:6(n − 3)4,7,10,13,16,19-Docosahexaenoic

Other double bond combinations or positions are possible as well.

Suitable fatty residues can furthermore be branched, for example, cancontain a methyl group in an iso or anteiso position of the fatty acidchain, or else closer to the chain middle, as in 10-R-methyloctadecanoicacid or tuberculostearic chain. Relatively important amongst branchedfatty acids are also isoprenoids, many of which are derived from3,7,11,15-tetramethylhexadec-trans-2-en-1-ol, the aliphatic alcoholmoiety of chlorophyll. Examples include5,9,13,17-tetramethyloctadecanoic acid and especially3,7,11,15-tetramethylhexadecanoic (phytanic) and2,6,10,14-tetramethylpentadecanoic (pristanic) acids. A good source of4,8,12-trimethyltridecanoic acid are marine organisms. Combination ofdouble bonds and side chains on a fatty residue are also possible.

Alternatively, suitable fatty residues may carry one or a few oxy- orcyclic groups, especially in the middle or towards the end of a chain.The most prominent amongst the later, alicyclic fatty acids, are thosecomprising a cyclopropane (and sometimes cyclopropane) ring, butcyclohexyl and cycloheptyl rings can also be found and might be usefulfor purposes of this disclosure. 2-(D)-Hydroxy fatty acids are moreubiquitous than alicyclic fatty acids, and are also importantconstituents of sphingolipids. Also interesting are15-hydroxy-hexadecanoic and 17-hydroxy-octadecanoic acids, and maybe9-hydroxy-octadeca-trans-10,trans-12-dienoic (dimorphecolic) and13-hydroxy-octadeca-cis-9,trans-11-dienoic (coriolic) acid. Arguably themost prominent hydroxyl-fatty acid in current pharmaceutical use isricinoleic acid, (D-(−)12-hydroxy-octadec-cis-9-enoic acid, whichcomprises up to 90% of castor oil, which is also often used inhydrogenated form. Epoxy-, methoxy-, and furanoid-fatty acids are ofonly limited practical interest in the context of this disclosure.

Generally speaking, unsaturation, branching or any other kind ofderivatization of a fatty acid is best compatible with the intention ofpresent disclosure of the site of such modification is in the middle orterminal part of a fatty acid chain. The cis-unsaturated fatty acids arealso more preferable than trans-unsaturated fatty acids and the fattyradicals with fewer double bonds are preferred over those with multipledouble bonds, due to oxidation sensitivity of the latter. Moreover,symmetric chain lipids are generally better suited than asymmetric chainlipids.

A preferred lipid of the Formula II is, for example, a naturalphosphatidylcholine, which used to be called lecithin. It can beobtained from egg (rich in palmitic, C_(16:0), and oleic, C_(18:1), butalso comprising stearic, C_(18:0), palmitoleic, C_(16:1), linolenic,C_(18:2), and arachidonic, C_(20:4), radicals), soybean (rich inunsaturated C₁₈ chains, but also containing some palmitic radical,amongst a few others), coconut (rich in saturated chains), olives (richin monounsaturated chains), saffron (safflower) and sunflowers (rich inn-6 linoleic acid), linseed (rich in n-3 linolenic acid), from whale fat(rich in monounsaturated n-3 chains), from primrose or primula (rich inn-3 chains). Preferred, natural phosphatidyl ethanolamines (used to becalled cephalins) frequently originate from egg or soybeans. Preferredsphingomyelins of biological origin are typically prepared from eggs orbrain tissue. Preferred phosphatidylserines also typically originatefrom brain material whereas phosphatidylglycerol is preferentiallyextracted from bacteria, such as E. Coli, or else prepared by way oftransphosphatidylation, using phospholipase D, starting with a naturalphosphatidylcholine. The preferably used phosphatidylinositols areisolated from commercial soybean phospholipids or bovine liver extracts.The preferred phosphatidic acid is either extracted from any of thementioned sources or prepared using phospholipase D from a suitablephosphatidylcholine.

Furthermore, synthetic phosphatidyl cholines (R⁴ in Formula IIcorresponds to 2-trimethylammonium ethyl), and R¹ and R² are aliphaticchains, as defined in the preceding paragraph with 12 to 30 carbonatoms. preferentially with 14 to 22 carbon atoms, and even morepreferred with 16 to 20 carbon atoms, under the proviso that the chainsmust be chosen so as to ensure that the resulting ESAs comprise fluidlipid bilayers. This typically means use of relatively short saturatedand of relatively longer unsaturated chains. Synthetic sphingomyelins(R⁴ in Formula IIB corresponds to 2-trimethylammonium ethyl), and R¹ isan aliphatic chain, as defined in the preceding paragraph, with 10 to 20carbon atoms, preferentially with 10 to 14 carbon atoms per fullysaturated chain and with 16-20 carbon atoms per unsaturated chain.

Synthetic phosphatidyl ethanolamines (R⁴ is 2-aminoethyl), syntheticphosphatidic acids (R⁴ is a proton) or its ester (R⁴ corresponds, forexample, to a short-chain alkyl, such as methyl or ethyl), syntheticphosphatidyl serines (R⁴ is L- or D-serine), or synthetic phosphatidyl(poly)alcohols, such as phosphatidyl inositol, phosphatidyl glycerol (R⁴is L- or D-glycerol) are preferred as lipids, wherein R¹ and R² arefatty residues of identical or moderately different type and length,especially such as given in the corresponding tables given before in thetext. Moreover, R¹ can represent alkenyl and R² identical hydroxyalkylgroups, such as tetradecylhydroxy or hexadecylhydroxy, for example, inditetradecyl or dihexadecylphosphatidyl choline or ethanolamine, R¹ canrepresent alkenyl and R² hydroxyacyl, such as a plasmalogen (R⁴trimethylammonium ethyl), or R¹ can be acyl, such as lauryl, myristoylor palmitoyl and R² can represent hydroxy as, for example, in natural orsynthetic lysophosphatidyl cholines or lysophosphatidyl glycerols orlysophosphatidyl ethanolamines, such as 1-myristoyl or1-palmitoyllysophosphatidyl choline or -phosphatidyl ethanolamine;frequently, R³ represents hydrogen.

A lipid of Formula IIB is also a suitable lipid within the sense of thisdisclosure. In Formula IIB, n=1, R¹ is an alkenyl group, R² is anacylamido group, R³ is hydrogen and R⁴ represents 2-trimethylammoniumethyl (choline group). Such a lipid is known under the name ofsphingomyelin.

Suitable lipids furthermore are a lysophosphatidyl choline analog, suchas 1-lauroyl-1,3-dihydroxypropane-3-phosphoryl choline, a monoglyceride,such as monoolein or monomyristin, a cerebroside, ceramide polyhexoside,sulfatide, sphingoplasmalogen, a ganglioside or a glyceride, which doesnot contain a free or esterified phosphoryl or phosphono or phosphinogroup in the 3 position. An example of such a glyceride isdiacylglyceride or 1-alkenyl-1-hydroxy-2-acyl glyceride with any acyl oralkenyl groups, wherein the 3-hydroxy group is etherified by one of thecarbohydrate groups named, for example, by a galactosyl group such as amonogalactosyl glycerin.

Lipids with desirable head or chain group properties can also be formedby biochemical means, for example, by means of phospholipases (such asphospholipase A1, A2, B, C and, in particular, D), desaturases,elongases, acyl transferases, etc., from natural or syntheticprecursors.

Furthermore, a suitable lipid is any lipid, which is contained inbiological membranes and can be extracted with the help of apolarorganic solvents, such as chloroform. Aside from the lipids alreadymentioned, such lipids also include, for example, steroids, such asestradiol, or sterols, such as cholesterol, beta-sitosterol,desmosterol, 7-keto-cholesterol or beta-cholestanol, fat-solublevitamins, such as retinoids, vitamins, such as vitamin A1 or A2, vitaminE, vitamin K, such as vitamin K1 or K2 or vitamin D1 or D3, etc.

The less soluble amphiphilic components comprise or preferably comprisea synthetic lipid, such as myristoleoyl, palmitoleoyl, petroselinyl,petroselaidyl, oleoyl, elaidyl, cis- or trans-vaccenoyl, linolyl,linolenyl, linolaidyl, octadecatetraenoyl, gondoyl, eicosaenoyl,eicosadienoyl, eicosatrienoyl, arachidoyl, cis- or trans-docosaenoyl,docosadienoyl, docosatrienoyl, docosatetraenoyl, lauroyl, tridecanoyl,myristoyl, pentadecanoyl, palmitoyl, heptadecanoyl, stearoyl ornonadecanoyl, glycerophospholipid or corresponding derivatives withbranched chains or a corresponding dialkyl or sphingosin derivative,glycolipid or other diacyl or dialkyl lipid.

The more soluble amphiphilic components(s) is/are frequently derivedfrom the less soluble components listed above and, to increase thesolubility, substituted and/or complexed and/or associated with abutanoyl, pentanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoylor undecanoyl substituent or several, mutually independent, selectedsubstituents or with a different material for improving the solubility.

A further suitable lipid is a diacyl- or dialkyl-glycerophosphoetha-nolamine azo polycthoxylene derivative, a didecanoylphosphatidyl cholineor a diacylphosphoolligomaltobionamide.

In certain embodiments, the amount of lipid in the formulation is fromabout 1% to about 12%, about 1% to about 10%, about 1% to about 4%,about 4% to about 7% or about 7% to about 10% by weight. In a specificembodiment, the lipid is a phospholipid. In another specific embodiment,the phospholipid is a phosphatidylcholine.

In some embodiments, the lipid in the formulation does not comprise analkyl-lysophospholipid. In some embodiments, the lipid in theformulation does not comprise a polyeneylphosphatidylcholine.

SURFACTANT

The term “surfactant” has its usual meaning. A list of relevantsurfactants and surfactant related definitions is provided in EP 0 475160 A1 (see, e.g., p. 6, 1. 5 to p.14. 1.17) and U.S. Pat. No. 6,165,500(see, e.g., col. 7, 1. 60 to col. 19,1. 64), each herein incorporated byreference in their entirety, and in appropriate surfactant orpharmaceutical Handbooks, such as Handbook of Industrial Surfactants orUS Pharmacopoeia, Pharm. Eu. In some embodiments, the surfactants arethose described in Tables 1-18 of U.S. Patent Application PublicationNo. 2002/0012680 A1, published Jan. 31, 2002, the disclosure of which isherein incorporated by reference in its entirety. The following listtherefore only offers a selection, which is by no means complete orexclusive, of several surfactant classes that are particularly common oruseful in conjunction with present patent application. Preferredsurfactants to be used in accordance with the disclosure include thosewith an HLB greater than 12. The list includes ionized long-chain fattyacids or long chain fatty alcohols, long chain fatty ammonium salts,such as alkyl- or alkenoyl-trimethyl-, -dimethyl- and -methyl-ammoniumsalts, alkyl- or alkenoyl-sulphate salts. long fatty chaindimethyl-aminoxides, such as alkyl- or alkenoyl-dimethyl-aminoxides,long fatty chain, for example alkanoyl, dimethyl-aminoxides andespecially dodecyl dimethyl-aminoxide, long fatty chain, for examplealkyl-N-methylglucamide- s and alkanoyl-N-methylglucamides, such asMEGA-8, MEGA-9 and MEGA-10, N-long fatty chain-N,N-dimethylglycines, forexample N-alkyl-N,N-dimethylglycines, 3-(long fattychain-dimethylammonio)-alkane-sulphonates, for example3-(acyidimethylammonio)-alkanesulphonates, long fatty chain derivativesof sulphosuccinate salts. such as bis(2-ethylalkyl) sulphosuccinatesalts, long fatty chain-sulphobetaines, for example acyl-sulphobetaines,long fatty chain betaines, such as EMPIGEN BB or ZWITTERGENT-3-16,-3-14, -3-12, -3-10, or -3-8, or polyethylen-glycol-acylphenyl ethers,especially nonaethylen-glycol-octyl- phenyl ether, polyethylene-longfatty chain-ethers, especially polyethylene-acyl ethers, such asnonaethylen-decyl ether, nonaethylen-dodecyl ether oroctaethylene-dodecyl ether, polyethyleneglycol-isoacyl ethers, such asoctaethyleneglycol-isotridecyl ether, polyethyleneglycol-sorbitane-longfatty chain esters, for example polyethyleneglycol-sorbitane-acyl estersand especially polyoxyethylene-monolaurate (e.g. polysorbate 20 or Tween20), polyoxyethylene-sorbitan-monooleate (e.g. polysorbate 80 or Tween80), polyoxyethylene-sorbitan-monolauroleylate,polyoxyethylene-sorbitan-monopetroselinate,polyoxyethylene-sorbitan-monoelaidate,polyoxyethylene-sorbitan-myristoleylate,polyoxyethylene-sorbitan-palmitoleinylate,polyoxyethylene-sorbitan-p-etroselinylate, polyhydroxyethylene-longfatty chain ethers, for example polyhydroxyethylene-acyl ethers, such aspolyhydroxyethylene-lauryl ethers, polyhydroxyethylene-myristoyl ethers,polyhydroxyethylene-cetylst-earyl, polyhyd roxyethylene-palmityl ethers,polyhyd roxyethylene-oleoyl ethers, polyhydroxyethylene-palmitoleoylethers, polyhydroxyethylene-lino- leyl, polyhydroxyethylen-4, or 6, or8, or 10, or 12-lauryl, miristoyl, palmitoyl, palmitoleyl, oleoyl orlinoeyl ethers (Brij series), or in the corresponding esters,polyhydroxyethylen-laurate, -myristate, -palmitate, -stearate or-oleate, especially polyhydroxyethylen-8-stearate (Myrj 45) andpolyhydroxyethylen-8-oleate, polyethoxylated castor oil 40 (CremophorEL), sorbitane-mono long fatty chain, for example alkylate (Arlacel orSpan series), especially as sorbitane-monolaurate (Arlacel 20, Span 20),long fatty chain, for example acyl-N-methylglucamides,alkanoyl-N-methylglucamides, especially decanoyl-N-methylglucamide,dodecanoyl-N-methylglucamide, long fatty chain sulphates, for examplealkyl-sulphates, alkyl sulphate salts, such as lauryl-sulphate (SDS),oleoyl-sulphate: long fatty chain thioglucosides, such asalkylthioglucosides and especially heptyl-, octyl- andnonyl-beta-D-thioglucopyranoside; long fatty chain derivatives ofvarious carbohydrates, such as pentoses, hexoses and disaccharides,especially alkyl-glucosides and maltosides, such as hexyl-, heptyl-,octyl-, nonyl- and decyl-beta-D-glucopyranoside or D-maltopyranoside;further a salt, especially a sodium salt, of cholate, deoxycholate,glycocholate, glycodeoxycholate, taurodeoxycholate, taurocholate, afatty acid salt, especially oleate, elaidate, linoleate, laurate, ormyristate, most often in sodium form, lysophospholipids,n-octadecylene-glycerophosphatidic acid,octadecylene-phosphorylglycerol, octadecylene-phosphorylserine, n-longfatty chain-glycero-phosphatidic acids, such asn-acyl-glycero-phosphatidic acids, especially laurylglycero-phosphatidic acids, oleoyl-glycero-phosphatidic acid, n-longfatty chain-phosphoryl glycerol, such as n-acyl-phosphorylglycerol,especially lauryl-, myristoyl-, oleoyl- orpalmitoeloyl-phosphorylglycerol, n-long fatty chain-phosphorylserine,such as n-acyl-phosphorylserine, especially lauryl-, myristoyl-, oleoyl-or palmitoeloyl-phosphorylserine, n-tetradecyl-glycero-phosphatidicacid, n-tetradecyl-phosphorylglycerol, n-tetradecyl-phosphorylserine,corresponding-, elaidoyl-, vaccenyl-lysophospholipids, correspondingshort-chain phospholipids, as well as all surface active and thusmembrane destabilising polypeptides. Surfactant chains are typicallychosen to be in a fluid state or at least to be compatible with themaintenance of fluid-chain state in carrier aggregates.

Table 5 lists preferred surfactants in accordance with one embodiment ofthe disclosure.

TABLE 5 Preferred surfactants Nonionic surfactants (S) Head/Type/TMFatty chain POE-sorbitan- POE-ether POE-ester POE-phenoxy- Length: nr.of ester Brij, Myrj, ether Selected Name(s) double bonds Tween MacrogolNonex Triton brandnames C24 Behen(o)yl C22 Eruca(o)yl C22:1-13cisArachin(o)yl C20 Gadolen(o)yl C20:1-11cis Arachidon(o)ylC20:4-5,8,11,14cis Ole(o)yl C18:1-9cis Tween 80 Brij 98 Simulsol-2599TritionX100** Stear(o)yl C18 Tween 60 Myrj-52 Linol(o)yl C18:2-9,12cisLinole(n/o)yl C18:3-9,12,15cis Palmitole(o)yl C18:1-9cis Palmit(o)yl C16Tween 40 NN Myrist(o)yl C14 Laur(o)yl C12 Tween 20 Brij 35 NN Capr(o)ylC10 Rel. concentration range L/S (M/M) 5/1-1/1 5/1-1/1 5/1-1/1 4/1-3/2NN: not readily available in the market but in principle suitable**Triton is not an oleate, but an octylphenoxy-POE derivative Myrj-45:Stearoyl-EO8; Myrj-49: Stearoyl-EO20 (not in the market); Myrj-59:Stearoyl-EO100; Myrj-52: Stearoyl-EO40; Simulsol-2599 =Macrogol-10-oleate Brij-98: Oleoyl-EO20 Brij-35: Lauryl-EO23

In certain embodiments, the surfactant is a nonionic surfactant. Thesurfactant may be present in the formulation in about 1% to about 10%,about 1% to about 4%, about 4% to about 7% or about 7% to about 10% byweight. In some embodiments, the amount of surfactants in theformulation is from about 0.2% to about 0.5%. In certain embodiments,the nonionic surfactant is selected from the group consisting of:polyoxyethylene sorbitans (polysobate surfactants), polyhydroxyethylenestearates or polyhydroxyethylene laurylethers (Brij surfactants). In aspecific embodiment, the surfactant is apolyoxyethylene-sorbitan-monooleate (e.g. polysorbate 80 or Tween 80).In certain embodiments, the polysorbate can have any chain with 12 to 20carbon atoms. In certain embodiments, the polysorbate is fluid in theformulation, which may contain one or more double bonds, branching, orcyclo-groups.

FORMULATIONS

In some embodiments, the formulations of the invention comprise only onelipid and only one surfactant. In other embodiments, the formulations ofthe invention comprise more than one lipid and only one surfactant,e.g., two, three, four, or more lipids and one surfactant. In otherembodiments, the formulations of the invention comprise only one lipidand more than one surfactant, e.g., two, three, four, or moresurfactants and one lipid. In other embodiments, the formulations of theinvention comprise more than one lipid and more than one surfactant,e.g., two, three, four, or more lipids and two, three, four, or moresurfactants.

The formulations of the invention may have a range of lipid tosurfactant ratios. The ratios may be expressed in terms of molar terms(mol lipid/mol surfactant). The molar ratio of lipid to surfactant inthe formulations may be from about 1:3 to about 30:1, from about 1:2 toabout 30:1, from about 1:1 to about 30:1, from about 5:1 to about 30:1,from about 10:1 to about 30:1, from about 15:1 to about 30:1, or fromabout 20:1 to about 30:1. In certain embodiments, the molar ratio oflipid to surfactant in the formulations of the invention may be fromabout 1:2 to about 10:1. In certain embodiments, the ratio is from about1:1 to about 2:1, from about 2:1 to about 3:1, from about 3:1 to about4:1, from about 4:1 to about 5:1 or from about 5:1 to about 10:1. Incertain embodiments, the molar ratio is from about 10.1 to about 30:1,from about 10:1 to about 20:1, from about 10:1 to about 25:1, and fromabout 20:1 to about 25:1. In specific embodiments, the lipid tosurfactant ratio is about 1.0:1.0, about 1.25:1.0, about 1.5/1.0, about1.75/1.0, about 2.0/1.0, about 2.5/1.0, about 3.0/1,0 or about 4.0/1.0.

The formulations of the invention may also have varying amounts of totalamount of the following components: lipid and surfactant combined (TA).The TA amount may be stated in terms of weight percent of the totalcomposition. In one embodiment, the TA is from about 1% to about 40%,about 5% to about 30%, about 7.5% to about 15%, about 5% to about 10%,about 10% to about 20% or about 20% to about 30%. In specificembodiments, the TA is 8%, 9%, 10%, 15% or 20%.

Selected ranges for total lipid amounts and lipid/surfactant ratios(mol/mol) for the formulations of the invention are described in Table 6below:

TABLE 6 Total Amount and Lipid to Surfactant Ratios TA (and surfactant)(%) Lipid/Surfactant (mol/mol) 5 to 10 1.0 to 1.25 5 to 10 1.25 to 1.755 to 10 1.75 to 2.25 5 to 10 2.25 to 3.00 5 to 10 3.00 to 4.00 5 to 104.00 to 8.00 5 to 10 10.00 to 13.00 5 to 10 15.00 to 20.00 5 to 10 20.00to 22.00 5 to 10 22.00 to 25.00 10 to 20 1.0 to 1.25 10 to 20 1.25 to1.75 10 to 20 1.25 to 1.75 10 to 20 2.25 to 3.00 10 to 20 3.00 to 4.0010 to 20 4.00 to 8.00 10 to 20 10.00 to 13.00 10 to 20 15.00 to 20.00 10to 20 20.00 to 22.00 10 to 20 22.00 to 25.00

The formulations of the invention do not comprise a pharmaceuticallyactive agent that has received marketing or regulatory approval in anycountry for the treatment of disorders related to fatty aciddeficiencies, fatty acid metabolism, hypertriglyceridemia andhypercholesterolemia, or any other disorder listed above.

The formulations of the invention may optionally contain one or more ofthe following ingredients: co-solvents, chelators, buffers,antioxidants, preservatives, microbicides, emollients, humectants,lubricants and thickeners. Preferred amounts of optional components aredescribed in Table 7.

The formulations of the invention may include a buffer to adjust the pHof the aqueous solution to a range from pH 3.5 to pH 9, pH 4 to pH 7.5,or pH 4 to pH 6.5. Examples of buffers include, but are not limited to,acetate buffers, lactate buffers, phosphate buffers, and propionatebuffers.

The formulations of the invention are typically formulated in aqueousmedia. The formulations may be formulated with or without co-solvents,such as lower alcohols

A “microbicide” or “antimicrobial” agent is commonly added to reduce thebacterial count in pharmaceutical formulations. Some examples ofmicrobicides are short chain alcohols, including ethyl and isopropylalcohol, chlorbutanol, benzyl alcohol, chlorbenzyl alcohol,dichlorbenzylalcohol, hexachlorophene; phenolic compounds, such ascresol, 4-chloro-m-cresol, p-chloro-m-xylenol, dichlorophene,hexachlorophene, povidon-iodine; parabenes, especially alkyl-parabenes,such as methyl-, ethyl-, propyl-, or butyl-paraben, benzyl paraben;acids, such as sorbic acid, benzoic acid and their salts; quaternaryammonium compounds, such as alkonium salts, e.g., a bromide,benzalkonium salts, such as a chloride or a bromide, cetrimonium salts,e.g., a bromide, phenoalkecinium salts, such as phenododecinium bromide,cetylpyridinium chloride and other salts; furthermore, mercurialcompounds, such as phenylmercuric acetate, borate, or nitrate,thiomersal, chlorhexidine or its gluconate, or any antibiotically activecompounds of biological origin, or any suitable mixture thereof.

Examples of “antioxidants” are butylated hydroxyanisol (BHA), butylatedhydroxytoluene (BHT) and di-tert-butylphenol (LY178002, LY256548,HWA-131, BF-389, CI-986, PD-127443, E-5119, BI-L-239XX, etc.), tertiarybutylhydroquinone (TBHQ), propyl gallate (PG),1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ); aromatic amines(diphenylamine, p-alkylthio-o-anisidine, ethylenediamine derivatives,carbazol, tetrahydroindenoindol); phenols and phenolic acids (guaiacol,hydroquinone, vanillin, gallic acids and their esters, protocatechuicacid, quinic acid, syringic acid, ellagic acid, salicylic acid,nordihydroguaiaretic acid (NDGA), eugenol); tocopherols (includingtocopherols (alpha, beta, gamma, delta) and their derivatives, such astocopheryl-acylate (e.g., -acetate, -laurate, myristate, -palmitate,-oleate, -linoleate, etc., or an y other suitable tocopheryl-lipoate),tocopheryl-POE-succinate; trolox and corresponding amide andthiocarboxamide analogues; ascorbic acid and its salts, isoascorbate, (2or 3 or 6)-o-alkylascorbic acids, ascorbyl esters (e.g., 6-o-lauroyl,myristoyl, palmitoyl-, oleoyl, or linoleoyl-L-ascorbic acid, etc.). Alsouseful are the preferentially oxidised compounds, such as sodiumbisulphite, sodium metabisulphite, thiourea; chellating agents, such asEDTA, GDTA, desferral; miscellaneous endogenous defence systems, such astransferrin, lactoferrin, ferritin, cearuloplasmin, haptoglobion,heamopexin, albumin, glucose, ubiquinol-10); enzymatic antioxidants,such as superoxide dismutase and metal complexes with a similaractivity, including catalase, glutathione peroxidase, and less complexmolecules, such as beta-carotene, bilirubin, uric acid; flavonoids(flavones, flavonols, flavonones, flavanonals, chacones, anthocyanins),N-acetylcystein, mesna, glutathione, thiohistidine derivatives,triazoles; tannines, cinnamic acid, hydroxycinnamatic acids and theiresters (coumaric acids and esters, caffeic acid and their esters,ferulic acid, (iso-) chlorogenic acid, sinapic acid); spice extracts(e.g., from clove, cinnamon, sage, rosemary, mace, oregano, allspice,nutmeg); carnosic acid, carnosol, carsolic acid; rosmarinic acid,rosmaridiphenol, gentisic acid, ferulic acid; oat flour extracts, suchas avenanthramide 1 and 2; thioethers, dithioethers, sulphoxides,tetralkylthiuram disulphides; phytic acid, steroid derivatives (e.g.,U74006F); tryptophan metabolites (e.g., 3-hydroxykynurenine,3-hydroxyanthranilic acid), and organochalcogenides.

“Thickeners” are used to increase the viscosity of pharmaceuticalformulations to and may be selected from selected from pharmaceuticallyacceptable hydrophilic polymers, such as partially etherified cellulosederivatives, comprising carboxymethyl-, hydroxyethyl-, hydroxypropyl-,hydroxypropylmethyl- or methyl-cellulose; completely synthetichydrophilic polymers comprising polyacrylates, polymethacrylates,poly(hydroxyethyl)-, poly(hydroxypropyl)-,poly(hydroxypropylmethyl)methacrylate, polyacrylonitrile,methallyl-sulphonate, polyethylenes, polyoxiethylenes, polyethyleneglycols, polyethylene glycol-lactide, polyethylene glycol-diacrylate,polyvinylpyrrolidone, polyvinyl alcohols, poly(propylmethacrylamide),poly(propylene fumarate-co-ethylene glycol), poloxamers,polyaspartamide, (hydrazine cross-linked) hyaluronic acid, silicone;natural gums comprising alginates, carrageenan, guar-gum, gelatine,tragacanth, (amidated) pectin, xanthan, chitosan collagen, agarose;mixtures and further derivatives or co-polymers thereof and/or otherpharmaceutically, or at least biologically, acceptable polymers.

The formulations of the present invention may also comprise a polarliquid medium. The formulations of the invention may be administered inan aqueous medium. The of the present invention may be in the form of asolution, suspension, emulsion, cream, lotion, ointment, gel, spray,film forming solution or lacquer.

In some embodiments, the invention relates to the use of a vesicularformulation as described above for the preparation of a pharmaceuticalcomposition for the treatment of disorders related to fatty aciddeficiencies, fatty acid metabolism, hypertriglyceridemia andhypercholesterolemia. In some embodiments, the invention relates to avesicular formulation or pharmaceutical composition comprising at leastone phospholipid and one nonionic surfactant for the treatment ofdisorders related to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia and hypercholesterolemia wherein the formulation orpharmaceutical composition is formulated for subcutaneous, topical orintravenous delivery.

Table 7 lists preferred excipients for the formulation.

TABLE 7 Preferred excipients for use in the formulations of theinvention Molar (M) Designated activity or Molar (M) Rel. Weight-Antioxidant or w %* Antibiotic % Buffer Molar Primary Butylatedhydroxyanisole, BHA 0.1-8 Acetate 30-150 mM Acetate 30-150 mM Butylatedhydroxytoluene, BHT 0.1-4 Benzyl alcohol 0.1-3 Phosphate  10-50 mMThymol 0.1-1 Butylparabene 0.1-3 Triethanolamine, HCL 30-150 mMMetabisulphite (MW = 190.1) 1-5 mM Ethylparabene 0.1-3 Bisulphite 1-5 mMImidurea (MW = 388.30) 0.1-1 Thiourea (MW = 76.12) 1-10 mM Dimethoxane(MW = 174.2  0.03-0.1 Monothioglycerol (MW = 108.16) 1-20 mMMethylparabene 0.1-5 Propyl gallate (MW = 212.2)  0.02-0.2Phenoxyethanol 0.1-5 Ascorbate (MW = 175.3 + ion) 1-10 mM Benzalkoniumchloride  0.01-0.2 Palmityl-ascorbate 0.01-1  Benzethonium chloride 0.01-0.1 Tocopherol-PEG 0.5-5 Phenol 0.05-2  Secondary (chelator)Phenylethyl alcohol 0.1-1 EDTA (MW = 292) 1-10 mM Thimerosal 0.005-0.1 EGTA (MW = 380.35) 1-10 mM Desferal (MW = 656.79) 0.1-5 mM *Aspercentage of Total Lipid quantity EGTA = Ethyleneglycol-bis-(2-aminoethyl)-N,N,N′,N′-tetraacetic acid EDTA =Ethylenedioxy-diethylene-dinitrilo-tetraacetic acid

VESICULAR FORMULATIONS

While not to be limited to any mechanism of action or any theory, theformulations of the invention may form vesicles or ESAs characterized bytheir adaptability, deformability, or penetrability.

The term vesicle or aggregate “adaptability” which governs the“tolerable surface curvature” is defined as the ability of a givenvesicle or aggregate to change easily its properties, such as shape,elongation ratio, and surface to volume ratio. The vesicles of thisinvention may be characterized by their ability to adjust theaggregates' shape and properties to the anisotropic stress caused bypore crossing. Sufficient adaptability implies that a vesicle or anaggregate can sustain different unidirectional forces or stress, such asone caused by pressure, without extensive fragmentation, which defines a“stable” aggregate. If an aggregate passes through a barrier fulfillingthis condition the terms “adaptability” and (shape) “deformability” plus“permeability” are essentially equivalent. A “barrier” in the context ofthis invention is (as in, for example, EP 0 475 160 and WO 98/17255) abody with through-extending narrow pores, such narrow pores having aradius which is at least 25% smaller than the radius of the ESAs(considered as spherical) before said ESAs permeate through such pores.

The term “narrow” used in connection with a pore implies that the poreradius is significantly, typically at least 25%, smaller than the radiusof the entity tested with regard to its ability to cross the pore. Thenecessary difference typically should be greater for the narrower pores.Using 25% limit is therefore quite suitable for >150 nm diameterwhereas >100% difference requirement is more appropriate for the smallersystems, e.g., with <50 nm diameter. For diameters around 20 nm,aggregate diameter difference of at least 200% is often required.

The term “semipermeable” used in connection with a barrier implies thata solution can cross transbarrier openings whereas a suspension ofnon-adaptable aggregates (large enough for the above definition of“narrow” pores to apply) cannot. Conventional lipid vesicles (liposomes)made from any common phosphatidylcholine in the gel lamellar phase orelse from any biological phosphatidylcholine/cholesterol 1/1 mol/molmixture or else comparably large oil droplets, all having the specifiedrelative diameter, are three examples for such non-adaptable aggregates.

The term “stable” means that the tested aggregates do not change theirdiameter spontaneously or under the transport related mechanical stress(e.g. during passage through a semipermeable barrier) unacceptably,which most often means only to a pharmaceutically acceptable degree. A20-40% change is normally considered acceptable; the halving or doublingof aggregate diameter is borderline and a greater change in diameter istypically unacceptable. Alternatively and very conveniently, the changein aggregate diameter resulting from pore crossing under pressure isused to assess system stability; the same criteria are then applied asfor “narrow” pores, mutatis mutandis. To obtain the correct value foraggregate diameter change, a correction for flux/vortex effects may benecessary. These procedures are described in greater detail in thepublications of the applicant in Cevc et. al., Biochim. Biophys. Acta2002; 1564:21-30.

Non-destructing passage of ultradeformable, mixed lipid aggregatesthrough narrow pores in a semi-permeable barrier is thus diagnostic ofhigh aggregate adaptability. If pore radius is two times smaller thanthe average aggregate radius the aggregate must change its shape andsurface-to-volume ratio at least 100% to pass without fragmentationthrough the barrier. An easy and reversible change in aggregate shapeinevitably implies high aggregate deformability and requires largesurface-to-volume ratio adaptation. A change in surface-to-volume ratioper se implies: a) high volume compressibility, e.g. in the case ofcompact droplets containing material other than, and immiscible with,the suspending fluid; b) high aggregate membrane permeability, e.g. inthe case of vesicles that are free to exchange fluid between inner andouter vesicle volume.

The vesicles or ESAs of the present invention have “adaptability” thatcan be assessed using the following method: 1) measure the flux (j_(a))of the aggregate or ESA suspension through a semi-permeable membrane(e.g., gravimetrically) for different transport-driving trans barrierpressures (Δp); 2) calculate the pressure dependence of barrierpenetratability P for the suspension by dividing each measured fluxvalue by the corresponding pressure value: P(Δp)=j_(a)(Δp)/Δp; 3)monitor the ratio of final and starting vesicle diameter 2r_(ves)(Δp)2_(ves,0) (e.g. by dynamic light scattering), wherein 2r_(ves)(Δp) is the vesicle diameter after semi-permeable barrier passagedriven by Δp and 2 r_(ves,0) is the starting vesicle diameter, and ifnecessary make corrections for the flow-effects; and 4) align both datasets P(Δp) vs. r_(ves)(Δp)/r_(ves,0) to determine the co-existence rangefor high aggregate adaptability and stability.

It is also useful, but not essential, to parameterize experimentalpenetratability data within the framework of Maxwell-approximation interms of the necessary pressure value p* and in terms of maximumpenetratability value P_(max). It is plausible to sum-up all thecontributions to a moving aggregate energy (deformation energy/ies,thermal energy, the shearing work, etc.) into a single, total energy.The equilibrium population density of aggregate's energetic levels thenmay be taken to correspond to Maxwell's distribution. All aggregateswith a total energy greater than the activation energy, E f E_(A), arefinally concluded to penetrate the barrier. The pore-crossingprobability for such aggregates is then given by the following formula,where e is dimensionless aggregate energy units of the activation energyE_(A):

${P(e)} = {1 - {{erf}\left( \sqrt{\frac{1}{e}} \right)} + {\sqrt{\frac{4}{\pi \; e}} \cdot {\exp \left\lbrack {- \frac{1}{e}} \right\rbrack}}}$

It is therefore plausible to represent barrier penetrability of a givensuspension as a function of transport driving pressure by the followingformula, where P_(max) is the maximum possible penetratability of agiven barrier (for the aggregates with zero transport resistance thispenetrability is identical to the penetrability of the suspending mediumflux), and p* is an adjustable parameter that describes the pressuresensitivity, and thus the transport resistance, of the tested system(for barriers with a fixed pore radius this sensitivity is a function ofaggregate properties solely; for non-interacting particles thesensitivity is dominated by aggregate adaptability, allowing to make theassumption: a_(a) proportional to 1/p*

${P(p)} = {p_{\max} \cdot \left\{ {1 - {{erf}\left( \sqrt{\frac{p^{*}}{p}} \right)} + {\sqrt{\frac{4p^{*}}{\pi \; p}} \cdot {\exp \left\lbrack {- \frac{p^{*}}{P}} \right\rbrack}}} \right\}}$

Other methods of testing deformability and adaptability which may beused to characterize the compositions of the invention are set forth,for example, in U.S. Patent Application Publication Nos. 2004/0071767and 2004/0105881, each herein incorporated by reference as if set forthherein in their entirety.

METHODS OF ADMINISTRATION/TREATMENT

In another embodiment, the invention provides methods of treatingdisorders related to fatty acid deficiencies, fatty acid metabolism,hypolididemia, hypertriglyceridemia and hypercholesterolemia comprisingadministering to a subject in need thereof a pharmaceutical compositioncomprising at least one phospholipid and one nonionic surfactant. Inanother embodiment, the invention provides methods of treating disordersrelated to fatty acid deficiencies, fatty acid metabolism,hypolididemia, hypertriglyceridemia and hypercholesterolemia comprisingadministering to a subject in need thereof a pharmaceutical compositionconsisting essentially of at least one phospholipid and one nonionicsurfactant, a pharmaceutically acceptable carrier, and optionallybuffers, antioxidants, preservatives, microbicides, antimicrobials,emollients, co-solvents, and/or thickeners. In another embodiment, theinvention provides methods of treating disorders related to fatty aciddeficiencies, fatty acid metabolism, hypolididemia, hypertriglyceridemiaand hypercholesterolemia comprising administering to a subject in needthereof a pharmaceutical composition consisting of at least onephospholipid and one nonionic surfactant, a pharmaceutically acceptablecarrier, and one or more of the following: buffers, antioxidants,preservatives, microbicides, antimicrobials, emollients, co-solvents,and thickeners.

In another embodiment, the invention provides methods of treatingdisorders related to inflammation, asthma, bronchospasm,atherothrombatic cardiovascular disorders, venous thrombatic disorders,pain, dysmenorrheal, hypercholesterolemia, hypertriglyceridemia, fattyacid metabolism, metal or other toxicity, Alzheimer's disease, gout ormacular degeneration, such as AMD, or fungal infection, comprisingadministering to a subject in need thereof a pharmaceutical compositioncomprising at least one phospholipid and one nonionic surfactant,wherein the pharmaceutical composition sequesters organic matter uponadministration. In another embodiment, the invention provides methods oftreating disorders related to inflammation, asthma, bronchospasm,atherothrombatic cardiovascular disorders, venous thrombatic disorders,pain, dysmenorrheal, hypercholesterolemia, hypertriglyceridemia, fattyacid metabolism, metal or other toxicity, Alzheimer's disease. gout ormacular degeneration, such as AMD, or fungal infection comprisingadministering to a subject in need thereof a pharmaceutical compositionconsisting essentially of at least one phospholipid and one nonionicsurfactant, a pharmaceutically acceptable carrier, and optionallybuffers, antioxidants, preservatives, microbicides, antimicrobials,emollients, co-solvents, and/or thickeners, wherein the pharmaceuticalcomposition sequesters organic matter upon administration. In anotherembodiment, the invention provides methods of treating disorders relatedto inflammation, asthma, bronchospasm, atherothrombatic cardiovasculardisorders, venous thrombatic disorders, pain, dysmenorrheal,hypercholesterolemia, hypertriglyceridemia, fatty acid metabolism, metalor other toxicity, Alzheimer's disease, gout or macular degeneration,such as AMD, or fungal infection comprising administering to a subjectin need thereof a pharmaceutical composition consisting of at least onephospholipid and one nonionic surfactant, a pharmaceutically acceptablecarrier, and one or more of the following: buffers, antioxidants,preservatives, microbicides, antimicrobials, emollients, co-solvents,and thickeners, wherein the pharmaceutical composition sequestersorganic matter upon administration.

PACKAGES

In another embodiment, the invention provides a pharmaceutical packageor kit comprising one or more containers filled with the formulation ofthe invention, and instructions for administration of the formulation toa patient or subject in need thereof for treating disorders related tofatty acid deficiencies, fatty acid metabolism, hypertriglyceridemia andhypercholesterolemia. In certain embodiments, the formulation comprisesone or more phospholipids and one or more surfactants. In certainembodiments, the formulation does not comprise a non-lipidnon-surfactant pharmaceutically active agent that has received marketingor regulatory approval in any country for the treatment of disordersrelated to fatty acid deficiencies, fatty acid metabolism,hypertriglyceridemia and hypercholesterolemia, or other disorder listedabove. In various embodiments, the container comprises a formulationformulated as a suspension, emulsion, gel, cream, lotion, spray, filmforming solution or lacquer. The invention provides packages or kitsthat can be used in any of the above-described methods.

EXAMPLES Example 1 Example Formulations

The following exemplary formulations for topical application may beprepared by the following procedure:

1. Organic Phase Production, Which Contains All Lipophilic Excipients

The organic phase is produced by weighing the lipid, the surfactant, anyadditional lipophilic excipients into suitable containers followed bymixing these components into anoptically isotropic phase which appearsas a clear solution. During mixing, the organic phase will be heated up,but temperature must not rise above 45° C.

2. Aqueous Phase Production

The aqueous phase is prepared by weighing the non-lipophilic componentsand water, which serves as solvent, into suitable containers and thenmixing these components into a clear solution. During mixing, thetemperature will be elevated to 40° C.

3. Production of a Concentrated Intermediate by Combination of BothPhases

The isotropic organic phase and the clear aqueous phase are combinedunder stirring in a suitable vessel. Before and during the combinationthe temperature of both phases must be kept between 35° C. and 45° C.The resulting intermediate is homogenised mechanically at 40° C. Beforestarting homogenisation, the pressure in the production vessel islowered to −0.08 MPa. The desired average carrier size is typicallyreached after 10 minutes of homogenisation.

Three process parameters must be controlled carefully during theproduction of the concentrated intermediate: temperature, homogenisercirculation velocity, and overall processing time.

4. Production of the Final Bulk Product by Mixing the ConcentratedIntermediate with Dilution Buffer.

The concentrated intermediate is diluted with the dilution buffer to theintended final concentration. The mixture is carefully stirred in themixing vessel at 20° C. to homogeneity.

Table 8 describes the amounts of surfactant and lipids, and otherexcipients in the transfersomes formulations, described in terms of thepercent of the total amount of formulation.

TABLE 8 Preferred Formulations Table 8A: This table lists the relativeamounts of each of the components of Preferred Formulations SurfactantLipid mg/g (1 to Buffer Antimicrobials Antioxidants Emollient OtherChelator mg/g 10% by wt.) (pH 4-7.5) (0-10 mg/g) (0-10 mg/g) (0-50 mg/g)(0-50 mg/g) (0-25 mg/g) 1 47.944 42.056 4 5.000 0.700 30.000 30.0003.000 2 53.750 31.250 4 5.000 0.700 30.000 15.000 3.000 3 90.561 79.4394 5.000 0.700 30.000 30.000 3.000 4 47.944 42.056 5 5.000 0.700 30.00030.000 3.000 5 50.607 44.393 5 5.000 0.700 0.000 10.000 3.000 6 90.56179.439 5 5.000 0.700 30.000 30.000 3.000 7 49.276 43.224 6.5 5.000 0.70030.000 30.000 3.000 8 53.750 31.250 6.5 5.000 0.200 30.000 0.000 3.000 990.561 79.439 6.5 5.000 0.200 30.000 20.000 3.000 10 41.351 48.649 45.000 0.200 30.000 30.000 3.000 11 47.882 37.118 4 5.000 0.200 0.00030.000 3.000 12 95.764 74.236 4 5.000 0.200 30.000 30.000 3.000 1365.676 24.324 5 5.000 0.200 0.000 25.000 3.000 14 62.027 22.973 5 5.0000.200 0.000 30.000 3.000 15 124.054 45.946 5 5.000 0.200 15.000 36.5103.000 16 62.687 32.313 6.5 5.000 0.200 15.000 0.000 3.000 17 41.85343.147 6.5 5.000 0.200 30.000 30.000 3.000 18 95.764 74.236 6.5 5.0000.200 0.000 30.000 3.000 19 47.882 37.118 6.5 5.000 0.200 0.000 0.0003.000 20 45.000 45.000 6.5 5.000 0.200 0.000 0.000 1.000 21 31.93558.065 5 5.000 0.200 30.000 15.000 3.000 22 42.500 42.500 6.5 5.0000.200 30.000 0.000 3.000 23 38.276 51.724 4 5.000 0.200 0.000 36.5103.000 24 42.500 42.500 4 5.000 0.200 0.000 15.000 3.000 25 85.000 85.0004 5.000 0.200 30.000 30.000 3.000 26 38.276 51.724 5 5.000 0.200 30.0000.000 1.000 27 36.429 48.571 5 5.000 0.200 30.000 30.000 3.000 28 72.29997.701 5 5.000 0.200 30.000 15.000 3.000 29 46.250 46.250 6.5 5.0000.700 0.000 20.000 3.000 30 38.804 46.196 6.5 5.000 0.700 15.000 30.0003.000 31 36.667 33.333 6.5 5.000 0.700 30.000 10.000 3.000 32 66.66723.333 4 5.000 0.200 0.000 0.000 3.000 33 45.833 41.667 4 5.000 0.20030.000 0.000 3.000 34 31.957 38.043 4 5.000 0.200 0.000 30.000 3.000 3547.143 42.857 5 5.000 0.200 30.000 25.000 1.000 36 96.905 88.095 5 5.0000.200 30.000 20.000 3.000 37 31.957 38.043 5 5.000 0.200 0.000 30.0003.000 38 35.455 54.545 6.5 5.000 0.700 30.000 0.000 3.000 39 84.457100.543 6.5 5.000 0.700 30.000 30.000 3.000 40 89.048 80.952 6.5 5.0000.700 30.000 30.000 3.000 41 41.087 48.913 4 5.000 0.700 30.000 36.5103.000 42 45.280 39.720 4 5.000 0.700 0.000 0.000 3.000 43 107.500 62.5004 5.000 0.700 30.000 30.000 3.000 44 77.243 67.757 4 5.000 0.700 0.00015.000 3.000 45 45.280 39.720 5 5.000 0.700 0.000 20.000 3.000 46 90.56179.439 5 5.000 0.700 0.000 30.000 3.000 47 47.944 42.056 5 5.000 0.7000.000 10.000 3.000 48 50.607 44.393 5.5 5.000 0.700 30.000 0.000 1.00049 107.500 62.500 5.5 5.000 0.700 30.000 0.000 3.000 50 47.944 42.0565.5 5.000 0.700 30.000 30.000 3.000 51 46.364 38.636 4 5.000 0.20030.000 25.000 3.000 52 46.364 38.636 4 5.000 0.200 0.000 20.000 3.000 5346.098 43.902 5 5.000 0.200 15.000 30.000 3.000 54 43.537 41.463 5 5.0000.200 30.000 0.000 3.000 55 45.000 45.000 5 5.000 0.200 0.000 30.0003.000 56 59.492 30.508 6.5 5.000 0.200 30.000 30.000 3.000 57 39.05445.946 6.5 5.000 0.200 0.000 0.000 3.000 58 35.854 34.146 6.5 5.0000.200 30.000 0.000 3.000 59 50.000 40.000 6.5 5.000 0.700 30.000 30.0003.000 60 38.571 51.429 6.5 5.000 0.700 30.000 30.000 3.000 61 41.95450.546 6.5 5.000 0.700 30.000 30.000 3.000 62 42.632 47.368 6.5 5.0000.700 30.000 30.000 3.000 63 46.098 43.902 6.5 5.000 0.700 30.000 30.0003.000 64 39.721 50.279 6.5 5.000 0.700 30.000 30.000 3.000 65 44.19850.802 6.5 5.000 0.700 30.000 30.000 3.000 66 46.453 51.047 6.5 5.0000.700 30.000 30.000 3.000 67 51.221 43.779 6.5 5.000 0.700 30.000 30.0003.000 68 54.167 43.333 6.5 5.000 0.700 30.000 30.000 3.000 69 66.44023.560 6.5 5.000 0.700 30.000 30.000 3.000 70 66.440 23.560 6.5 5.0000.700 30.000 30.000 3.000 71 66.440 23.560 6.5 5.000 0.700 30.000 30.0003.000 72 40.000 50.000 6.5 5.000 0.700 30.000 30.000 3.000 73 40.00050.000 6.5 5.000 0.700 30.000 30.000 3.000 74 40.000 50.000 5.5 0.0000.700 30.000 30.000 3.000 75 40.000 50.000 6.5 5.000 0.700 30.000 30.0003.000 76 40.000 50.000 6.5 5.000 0.700 30.000 30.000 3.000 77 40.00050.000 6.5 5.000 0.700 30.000 30.000 3.000 78 66.440 23.560 6.5 5.0000.700 30.000 30.000 3.000 79 66.440 23.560 6.5 5.000 0.700 30.000 30.0003.000 80 40.000 50.000 5.5 0.000 0.700 30.000 30.000 3.000 81 40.00050.000 5.5 5.000 0.700 30.000 30.000 3.000 82 44.444 55.556 5.5 5.0000.700 30.000 30.000 3.000 83 66.440 23.560 5.5 5.000 0.700 30.000 30.0003.000 84 54.000 36.000 4 5.000 0.700 30.000 30.000 3.000 85 50.00040.000 4 5.000 0.700 30.000 30.000 3.000 86 48.611 38.889 4 5.000 0.70030.000 30.000 3.000 87 46.575 38.425 4 5.000 0.700 30.000 30.000 3.00088 46.575 38.425 4 5.000 0.700 30.000 30.000 3.000 89 46.575 38.425 45.000 0.700 30.000 30.000 3.000 90 50.000 40.000 4.5 5.000 0.700 30.00030.000 3.000 91 94.444 75.556 4 5.000 0.700 30.000 30.000 3.000 9246.712 38.288 4 5.000 0.700 30.000 30.000 3.000 93 48.889 39.111 4 5.0000.700 30.000 30.000 3.000 94 39.721 50.279 6.5 5.000 0.700 30.000 30.0003.000 95 90.000 0.000 6.5 5.000 0.700 30.000 30.000 3.000 96 68.7008.500 7.5 5.000 0.700 30.000 36.510 1.000 97 71.460 4.720 7.8 5.0000.700 50.000 35.000 3.000 99 71.460 4.720 7.8 5.000 0.700 50.000 15.0003.000 98 71.460 4.720 7.8 0.000 0.700 50.000 15.000 3.000 100 71.4604.720 7.8 0.000 0.700 50.000 35.000 3.000 101 46.575 38.425 4 0.0000.700 0.000 0.000 3.000 102 46.575 38.425 4 0.000 0.700 0.000 0.0003.000 103 54.643 30.357 4 5.000 0.700 0.000 0.000 3.000 104 39.72 50.2796.5 5.000 0.700 30.000 30.000 3.000 105 90.00 6.5 5.000 0.700 30.00030.000 3.000 106 46.57 38.425 4 0.700 3.000 107 46.75 38.425 4 0.7003.000 108 54.64 30.357 4 0.700 3.000 109 46.364 38.636 4 5.000 0.20030.000 25.000 3.000 110 46.364 38.636 4 5.000 0.200 0.000 20.000 3.000111 46.098 43.902 5 5.000 0.200 15.000 30.000 3.000 112 43.537 41.463 55.000 0.200 30.000 0.000 3.000 113 45.000 45.000 5 5.000 0.200 0.00030.000 3.000 114 59.492 30.508 6.5 5.000 0.200 30.000 30.000 3.000 11539.054 45.946 6.5 5.000 0.200 0.000 0.000 3.000 116 35.854 34.146 6.55.000 0.200 30.000 0.000 3.000 117 50.000 40.000 6.5 5.000 0.700 30.00030.000 3.000 118 38.571 51.429 6.5 5.000 0.700 30.000 30.000 3.000 11941.954 50.546 6.5 5.000 0.700 30.000 30.000 3.000 120 42.632 47.368 6.55.000 0.700 30.000 30.000 3.000 121 46.098 43.902 6.5 5.000 0.700 30.00030.000 3.000 122 39.721 50.279 6.5 5.000 0.700 30.000 30.000 3.000 12344.198 50.802 6.5 5.000 0.700 30.000 30.000 3.000 124 46.453 51.047 6.55.000 0.700 30.000 30.000 3.000 125 51.221 43.779 6.5 5.000 0.700 30.00030.000 3.000 126 54.167 43.333 6.5 5.000 0.700 30.000 30.000 3.000 12766.440 23.560 6.5 5.000 0.700 30.000 30.000 3.000 128 66.440 23.560 6.55.000 0.700 30.000 30.000 3.000 129 66.440 23.560 6.5 5.000 0.700 30.00030.000 3.000 Table 8B: The table lists the specific components of theformulas listed above. Formula Lipid Surfactant Buffer AntimicrobialAntioxidants Emollient Chelator Other 1-4 Sphingomyelin, e.g., Tween 80Lactate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol brain orparaben sodium metabisulfite (0.500) 5-7 Sphingomyelin, Brij 98 AcetateBenzyl alcohol BHT (0.200) Glycerol EDTA Ethanol lauroyl or parabensodium metabisulfite (0.500)  8-12 Phosphatidyl choline + Brij 98Phosphate Benzyl alcohol HTHQ Glycerol EDTA Ethanol Phosphatidylglycerol or paraben 13-16 Phosphatidyl choline + Span 20 Acetate Benzylalcohol HTHQ Glycerol EDTA Ethanol phosphatidylinositol or paraben 17-18Phosphatidyl choline + Tween 80 Phosphate Benzyl alcohol BHT GlycerolEDTA Ethanol phosphatidic acid or paraben 19 Phosphatidyl choline + Brij98 + Phosphate Benzyl alcohol BHT Glycerol EDTA Ethanol phosphatidicacid Tween 80 or paraben 20 Phosphatidyl choline + Span 20 + PhosphateBenzyl alcohol BHT Glycerol EDTA Ethanol phosphatidic acid Tween 80 orparaben 21 Phosphatidyl choline Cremophor + Lactate Thimerosal BHAGlycerol EDTA Ethanol Span 20 22 Phosphatidyl choline Cremophor +Lactate Thimerosal BHA Glycerol EDTA Ethanol Tween 80 23-28 Phosphatidylcholine Cremophor Lactate Thimerosal BHA Glycerol EDTA Ethanol 29-30Phosphatidyl Tween 80 Phosphate Thimerosal BHT (0.200) Glycerol EDTAEthanol ethanolamine sodium metabisulfite (0.500) 31 Phosphatidyl Brij98 + Phosphate Thimerosal BHT (0.200) Glycerol EDTA Ethanol ethanolamineTween 80 sodium metabisulfite (0.500) 32 Phosphatidyl glycerolCremophor + Acetate Benzyl alcohol BHT Glycerol EDTA Ethanol Brij 98 orparaben 33-37 Phosphatidyl glycerol Brij 98 Acetate Benzyl alcohol BHTGlycerol EDTA Ethanol or paraben 38-40 Phosphatidyl Cremophor phosphateBenzyl alcohol BHT (0.200) Glycerol EDTA Ethanol ethanolamine or parabensodium metabisulfite (0.500) 41-47 Phosphatidyl glycerol Tween 80Propionate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol or parabensodium metabisulfite (0.500) 48-50 Phosphatidyl serine Brij 98 PhosphateThimerosal BHT (0.200) Glycerol EDTA Ethanol sodium metabisulfite(0.500) 51-58 Phosphatidyl glycerol Brij 98 Acetate Benzyl alcohol BHTGlycerol EDTA Ethanol or paraben 59-68 Phosphatidyl choline Tween 80Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol or parabensodium metabisulfite (0.500) 69-71 Phosphatidyl choline Brij 98Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol or parabensodium metabisulfite (0.500) 72-73 Phosphatidyl choline Tween 80Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol or parabensodium metabisulfite (0.500) 74 Phosphatidyl choline Tween 80 AcetateBHT (0.200) Glycerol EDTA Ethanol sodium metabisulfite (0.500) 75Phosphatidyl choline Tween 80 Phosphate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 76Phosphatidyl choline Brij 98 Phosphate Benzalkonium BHT (0.200) GlycerolEDTA Ethanol chloride sodium metabisulfite (0.500) 77 Phosphatidylcholine Tween 80 Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTAEthanol or paraben sodium metabisulfite (0.500) 78 Phosphatidyl cholineBrij 98 Phosphate Benzalkonium BHT (0.200) Glycerol EDTA Ethanolchloride sodium metabisulfite (0.500) 79 Phosphatidyl choline Brij 98Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTA Ethanol or parabensodium metabisulfite (0.500) 80 Phosphatidyl choline Tween 80 AcetateBHT (0.200) Glycerol EDTA Ethanol sodium metabisulfite (0.500) 81Phosphatidyl choline Tween 80 Acetate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 82-83Phosphatidyl choline Tween 80 Acetate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 84-88Phosphatidyl choline Tween 80 Acetate Benzyl alcohol BHA (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 89Phosphatidyl choline Tween 80 Acetate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 90-93Phosphatidyl choline Tween 80 Acetate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 94-96Phosphatidyl choline Tween 80 Phosphate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 97-98Phosphatidyl choline Tween 80 Phosphate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500)  99-100Phosphatidyl choline Tween 80 Phosphate BHT (0.200) Glycerol EDTAEthanol sodium metabisulfite (0.500) 101-103 Phosphatidyl choline Tween80 Acetate BHT (0.200) EDTA sodium metabisulfite (0.500) 104 Phosphatidyl choline Tween 80 Phosphate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol or paraben sodium metabisulfite (0.500) 105 Phosphatidyl choline Phosphate Benzyl alcohol BHT (0.200) Glycerol EDTAEthanol or paraben sodium metabisulfite (0.500) 106-108 Phosphatidylcholine Tween 80 Phosphate BHT (0.200) EDTA sodium metabisulfite (0.500)109-116 Phosphatidyl glycerol Brij 98 Acetate Benzyl alcohol BHTGlycerol EDTA Ethanol and lysophospholipid or paraben 117-126Phosphatidyl choline Tween 80 Phosphate Benzyl alcohol BHT (0.200)Glycerol EDTA Ethanol and lysophospholipid or paraben sodiummetabisulfite (0.500) 127-129 Phosphatidyl choline Brij 98 PhosphateBenzyl alcohol BHT (0.200) Glycerol EDTA Ethanol and lysophospholipid orparaben sodium metabisulfite (0.500)

Example Formulation 1

Formulation 1 comprises sphingomyelin (brain) (47.944 mg/g) as a lipid,Tween 80 (42.056mg/g) as a surfactant, lactate buffer (pH 4). benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.0500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 2

Formulation 2 comprises sphingomyelin (brain) (53.750 mg/g) as a lipid,Tween 80 (31.250 mg/g) as a surfactant, lactate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(15.000 mg/g).

Example Formulation 3

Formulation 3 comprises sphingomyelin (brain) (90.561 mg/g) as a lipid,Tween 80 (79.439 mg/g) as a surfactant, lactate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 4

Formulation 4 comprises sphingomyelin (brain) (47.944 mg/g) as a lipid,Tween 80 (42.056 mg/g) as a surfactant, lactate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 5

Formulation 5 comprises sphingomyelin lauroyl (50.607 mg/g) as a lipid,Brij 98 (44.393 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, EDTA (3.000mg/g) as a chelating agent, and ethanol (10.000 mg/g).

Example Formulation 6

Formulation 6 comprises sphingomyelin lauroyl (90.561 mg/g) as a lipid,Brij 98 (79.439 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 meg) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 7

Formulation 7 comprises sphingomyelin lauroyl (49.276 mg/g) as a lipid,Brij 98 (79.439 mg/g) as a surfactant, acetate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 8

Formulation 8 comprises phosphatidyl choline and phosphatidyl glycerol(53.750 mg/g) as a lipid, Brij 98 (31.250 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), and EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 9

Formulation 9 comprises phosphatidyl choline and phosphatidyl glycerol(90.561 mg/g) as a lipid, Brij 98 (79.439 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 10

Formulation 10 comprises phosphatidyl choline and phosphatidyl glycerol(41.351 mg/g) as a lipid, Brij 98 (48.649 mg/g) as a surfactant,phosphate (pH 4) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTIIQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 11

Formulation 11 comprises phosphatidyl choline and phosphatidyl glycerol(47.882 mg/g) as a lipid, Brij 98 (37.118 mg/g) as a surfactant,phosphate (pH 4) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol, EDTA(3.000 mg/g) as a chelating agent, and ethanol (30.000 mg/g).

Example Formulation 12

Formulation 12 comprises phosphatidyl choline and phosphatidyl glycerol(95.764 mg/g) as a lipid, Brij 98 (74.236 mg/g) as a surfactant,phosphate (pH 4) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 13

Formulation 13 comprises phosphatidyl choline and phosphatidylinositol(66.676 mg/g) as a lipid, Span 20 (24.324 mg/g) as a surfactant, acetate(pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g), HTHQ (0.200 mg/g)as an antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(25.000 mg/g).

Example Formulation 14

Formulation 14 comprises phosphatidyl choline and phosphatidylinositol(62.027 mg/g) as a lipid, Span 20 (22.973 mg/g) as a surfactant, acetate(pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, EDTA (3.000mg/g) as a chelating agent, and ethanol (30.000 mg/g).

Example Formulation 15

Formulation 15 comprises phosphatidyl choline and phosphatidylinositol(124.054 mg/g) as a lipid, Span 20 (45.946 mg/g) as a surfactant,acetate (pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), and EDTA (3.000 mg/g) as a chelating agent, and ethanol(36.510 mg/g).

Example Formulation 16

Formulation 16 comprises phosphatidyl choline and phosphatidylinositol(62.687 mg/g) as a lipid, Span 20 (32.313 mg/g) as a surfactant, acetate(pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, HTHQ (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 17

Formulation 17 comprises phosphatidyl choline and phosphatidic acid(41.853 mg/g) as a lipid, Tween 80 (43.147 mg/g) as a surfactant.phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, BHT (0.200 mg/g) as an antioxidant, glycerol(30.000 mg/g), EDTA (3.000 mg/g), and ethanol (30.000 mg/g).

Example Formulation 18

Formulation 18 comprises phosphatidyl choline and phosphatidic acid(95.764 mg/g) as a lipid, Tween 80 (74.236 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial agent, BHT (0.200 mg/g) as an antioxidant, EDTA (3.000mg/g), and ethanol (30.000 mg/g).

Example Formulation 19

Formulation 19 comprises phosphatidyl choline and phosphatidic acid(47.882 mg/g) as a lipid, Brij 98 and Tween 80 (37.118 mg/g) as asurfactant, phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000mg/g) as an antimicrobial agent, BHT (0.200 mg/g) as an antioxidant, andEDTA (3.000 mg/g).

Example Formulation 20

Formulation 20 comprises phosphatidyl choline and phosphatidic acid(45.000 mg/g) as a lipid, Span 20 and Tween 80 (45.000 mg/g) as asurfactant, phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000mg/g) as an antimicrobial agent, BHT (0.200 mg/g) as an antioxidant, andEDTA (1.000 mg/g).

Example Formulation 21

Formulation 21 comprises phosphatidyl choline (31.935 mg/g) as a lipid,cremophor and Span 20 (58.065 mg/g) as a surfactant, lactate (pH 5)buffer, thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200mg/g) as an antioxidant, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as achelating agent, and ethanol (15.000 mg/g).

Example Formulation 22

Formulation 22 comprises phosphatidyl choline (42.500 mg/g) as a lipid,cremophor and Tween 80 (42.500 mg/g) as a surfactant, lactate (pH 6.5)buffer, thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200mg/g) as an antioxidant, glycerol (30.000 mg/g), and EDTA (3.000 mg/g)as a chelating agent.

Example Formulation 23

Formulation 23 comprises phosphatidyl choline (38.276 mg/g) as a lipid,cremophor (51.724 mg/g) as a surfactant, lactate (pH 4) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(36.510 mg/g).

Example Formulation 24

Formulation 24 comprises phosphatidyl choline (42.500 mg/g) as a lipid,cremophor (42.500 mg/g) as a surfactant, lactate (pH 4) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(15.000 mg/g).

Example Formulation 25

Formulation 25 comprises phosphatidyl choline (85.000 mg/g) as a lipid,cremophor (85.000 mg/g) as a surfactant, lactate (pH 4) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 26

Formulation 26 comprises phosphatidyl choline (38.276 mg/g) as a lipid,cremophor (51.276 mg/g) as a surfactant, lactate (pH 5) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, and EDTA (1.000 mg/g) as a chelating agent.

Example Formulation 27

Formulation 27 comprises phosphatidyl choline (36.429 mg/g) as a lipid,cremophor (48.571 mg/g) as a surfactant, lactate (pH 5) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 28

Formulation 28 comprises phosphatidyl choline (72.299 mg/g) as a lipid,cremophor (97.701 mg/g) as a surfactant, lactate (pH 5) buffer,thimerosal (5.000 mg/g) as an antimicrobial agent, BHA (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(15.000 mg/g).

Example Formulation 29

Formulation 29 comprises phosphatidyl ethanolamine (46.250 mg/g) as alipid, Tween 80 (46.250 mg/g) as a surfactant, phosphate (pH 6.5)buffer, thimerosal (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as an antioxidant, EDTA(3.000 mg/g) as a chelating agent, and ethanol (20.000 mg/g).

Example Formulation 30

Formulation 30 comprises phosphatidyl ethanolamine (38.804 mg/g) as alipid, Tween 80 (46.196 mg/g) as a surfactant, phosphate (pH 6.5)buffer, thimerosal (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as an antioxidant, glycerol(15.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 31

Formulation 31 comprises phosphatidyl ethanolamine (3 6.667 mg/g) as alipid, Brij 98 and Tween 80 (33.333 mg/g) as a surfactant, phosphate (pH6.5) buffer, thimerosal (5.000 mg/g) as an antimicrobial agent, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (30.000 mg/g).

Example Formulation 32

Formulation 32 comprises phosphatidyl glycerol (23.333 mg/g) as a lipid,cremophor and Brij 98 (66.667 mg/g) as a surfactant, acetate (pH 4)buffer, benzyl alcohol or paraben (5.000 mg/g) as an antimicrobialagent, BHT (0.200 mg/g) as an antioxidant, and EDTA (3.000 mg/g) as achelating agent.

Example Formulation 33

Formulation 33 comprises phosphatidyl glycerol (45.833 mg/g) as a lipid,Brij 98 (41.667 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) as an antioxidant, glycerol (30.000 mg/g), and EDTA (3.000 mg/g)as a chelating agent.

Example Formulation 34

Formulation 34 comprises phosphatidyl glycerol (31.957 mg/g) as a lipid,Brij 98 (38.043 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) as an antioxidant, EDTA (3.000 mg/g) as a chelating agent, andethanol (30.000 mg/g).

Example Formulation 35

Formulation 35 comprises phosphatidyl glycerol (47.143 mg/g) as a lipid,Brij 98 (42.857 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) as an antioxidant, glycerol (30.000 mg/g), EDTA (1.000 mg/g) as achelating agent, and ethanol (25.000 mg/g).

Example Formulation 36

Formulation 36 comprises phosphatidyl glycerol (96.905 mg/g) as a lipid,Brij 98 (88.095 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) as an antioxidant, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as achelating agent, and ethanol (20.000 mg/g).

Example Formulation 37

Formulation 37 comprises phosphatidyl glycerol (31.957 mg/g) as a lipid,Brij 98 (38.043) as a surfactant, acetate (pH 5) buffer, benzyl alcoholor paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 38

Formulation 38 comprises phosphatidyl ethanolamine (35.455 mg/g) as alipid, cremophor (54.545 mg/g) as a surfactant, phosphate (pH 6.5)buffer, benzyl alcohol or paraben (5.000 mg/g) as an antimicrobialagent, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), and EDTA (3.000 mg/g) as achelating agent.

Example Formulation 39

Formulation 39 comprises phosphatidyl ethanolamine (84.457 mg/g) as alipid, cremophor (100.543 mg/g) as a surfactant, phosphate (pH 6.5)buffer, benzyl alcohol or paraben (5.000 mg/g) as an antimicrobialagent, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 40

Formulation 40 comprises phosphatidyl ethanolamine (89.048 mg/g) as alipid, cremophor (80.952 mg/g) as a surfactant, phosphate (pH 6.5)buffer, benzyl alcohol or paraben (5.000 mg/g), BHT (0.200 mg/g) andsodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 41

Formulation 41 comprises phosphatidyl glycerol (41.087 mg/g) as a lipid,Tween 80 (48.913 mg/g) as a surfactant, propionate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(36.510 mg/g).

Example Formulation 42

Formulation 42 comprises phosphatidyl glycerol (45.280 mg/g) as a lipid,Tween 80 (39.720 mg/g) as a surfactant, propionate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g), and EDTA (3.000 mg/g) as achelating agent.

Example Formulation 43

Formulation 43 comprises phosphatidyl glycerol (107.500 mg/g) as alipid, Tween 80 (62.500 mg/g) as a surfactant, propionate (pH 4) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (30.000 mg/g).

Example Formulation 44

Formulation 44 comprises phosphatidyl glycerol (77.243 mg/g) as a lipid,Tween 80 (67.757 mg/g) as a surfactant, propionate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants. EDTA (3.000mg/g) as a chelating agent, and ethanol (30.000 mg/g).

Example Formulation 45

Formulation 45 comprises phosphatidyl glycerol (45.280 mg/g) as a lipid,Tween 80 (39.720 mg/g) as a surfactant, propionate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, EDTA (3.000mg/g) as a chelating agent, and ethanol (30.000 mg/g).

Example Formulation 46

Formulation 46 comprises phosphatidyl glycerol (90.561 mg/g) as a lipid,Tween 80 (79.439 mg/g) as a surfactant, propionate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, EDTA (3.000mg/g) as a chelating agent, and ethanol (30.000 mg/g).

Example Formulation 47

Formulation 47 comprises phosphatidyl glycerol (47.944 mg/g) as a lipid,Tween 80 (42.056 mg/g) as a surfactant, propionate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial agent, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, EDTA (3.000mg/g) as a chelating agent, and ethanol (10.000 mg/g).

Example Formulation 48

Formulation 48 comprises phosphatidyl serine (50.607 mg/g) as a lipid,Brij 98 (44.393 mg/g) as a surfactant, phosphate (pH 5.5) buffer,thimerasol (5.000 mg/g) as an antimicrobial agent, BHT (0.200 mg/g) andsodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), and EDTA (1.000 mg/g) as a chelating agent.

Example Formulation 49

Formulation 49 comprises phosphatidyl serine (107.500 mg/g) as a lipid,Brij 98 (62.500 mg/g) as a surfactant, phosphate (pH 5.5) buffer,thimerasol (5.000 mg/g) as an antimicrobial agent, BHT (0.200 mg/g) andsodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), and EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 50

Formulation 50 comprises phosphatidyl serine (47.944 mg/g) as a lipid,Brij 98 (42.056 mg/g) as a surfactant, phosphate (pH 5.5) buffer,thimerasol (5.000 mg/g) as an antimicrobial agent, BHT (0.200 mg/g) andsodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 51

Formulation 51 comprises phosphatidyl glycerol (46.364 mg/g) as a lipid,Brij 98 (38.636 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (25.000 mg/g).

Example Formulation 52

Formulation 52 comprises phosphatidyl glycerol (46.364 mg/g) as a lipid,Brij 98 (38.636 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(20.000 mg/g).

Example Formulation 53

Formulation 53 comprises phosphatidyl glycerol (46.098 mg/g) as a lipid,Brij 98 (43.902 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or parabcn (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, glycerol (15.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 54

Formulation 54 comprises phosphatidyl glycerol (43.537 mg/g) as a lipid,Brij 98 (41.463 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, glycerol (30.000 mg/g), and EDTA (3.000 mg/g) as achelating agent.

Example Formulation 55

Formulation 55 comprises phosphatidyl glycerol (45.000 mg/g) as a lipid,Brij 98 (45.000 mg/g) as a surfactant, acetate (pH 5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 56

Formulation 56 comprises phosphatidyl glycerol (59.492 mg/g) as a lipid,Brij 98 (30.508 mg/g) as a surfactant, acetate (pH 6.5) buffer. benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 57

Formulation 57 comprises phosphatidyl glycerol (39.054 mg/g) as a lipid,Brij 98 (45.946 mg/g) as a surfactant, acetate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, and EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 58

Formulation 58 comprises phosphatidyl glycerol (35.854 mg/g) as a lipid,Brij 98 (34.146 mg/g) as a surfactant, acetate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g) asan antioxidant, glycerol (30.000 mg/g), and EDTA (3.000 mg/g) as achelating agent.

Example Formulation 59

Formulation 59 comprises phosphatidyl choline (50.000 mg/g) as a lipid,Tween 80 (40.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 60

Formulation 60 comprises phosphatidyl choline (38.571 mg/g) as a lipid,Tween 80 (51.429 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g), and ethanol (30.000 mg/g).

Example Formulation 61

Formulation 61 comprises phosphatidyl choline (41.954 mg/g) asphospholipid, Tween 80 (50.546 mg/g) as surfactant, phosphate (pH 6.5)buffer, benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (30.000 mg/g), EDTA (3.000 mg/g), and ethanol (30.000 mg/g).

Example Formulation 62

Formulation 62 comprises phosphatidyl choline (42.632 mg/g) as a lipid,Tween 80 (47.368 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200meg) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 63

Formulation 63 comprises phosphatidyl choline (46.098 mg/g) as a lipid,Tween 80 (43.902 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 64

Formulation 64 comprises phosphatidyl choline (39.721 mg/g) as a lipid,Tween 80 (50.279 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 65

Formulation 65 comprises phosphatidyl choline (44.198 mg/g) as a lipid,Tween 80 (50.802 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 66

Formulation 66 comprises phosphatidyl choline (46.453 mg/g) as a lipid,Tween 80 (51.047 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 67

Formulation 67 comprises phosphatidyl choline (51.221 mg/g) as a lipid,Tween 80 (43.779 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent. and ethanol(30.000 mg/g).

Example Formulation 68

Formulation 68 comprises phosphatidyl choline (54.167 mg/g) as a lipid,Tween 80 (43.333 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 69

Formulation 69 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Brij 98 (23.560 mg/g) as a surfactant, phosphate (pH 6,5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g). Example formulation 69 is an emulsion.

Example Formulation 70

Formulation 70 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Brij 98 (23.560 mg/g) as a surfactant, phosphate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g). Example formulation 70 is a suspension.

Example Formulation 71

Formulation 71 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Brij 98 (23.560 mg/g) as a surfactant, phosphate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0,500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 72

Formulation 72 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g). Example formulation 72 is an emulsion.

Example Formulation 73

Formulation 73 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g). Example formulation 73 is a suspension.

Example Formulation 74

Formulation 74 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, acetate (pH 5.5) buffer, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (30.000 mg/g).

Example Formulation 75

Formulation 75 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 76

Formulation 76 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Brij 98 (50.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzalkonium chloride (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 77

Formulation 77 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 78

Formulation 78 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Brij 98 (23.560 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzalkonium chloride (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 79

Formulation 79 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Brij 98 (23.560 mg/g) as a surfactant, phosphate (pH 6.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 80

Formulation 80 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, acetate (pH 5.5) buffer, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (30.000 mg/g).

Example Formulation 81

Formulation 81 comprises phosphatidyl choline (40.000 mg/g) as a lipid,Tween 80 (50.000 mg/g) as a surfactant, acetate (pH 5.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 82

Formulation 82 comprises phosphatidyl choline (44.444 mg/g) as a lipid,Tween 80 (55.556 mg/g) as a surfactant, acetate (pH 5.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 83

Formulation 83 comprises phosphatidyl choline (66.440 mg/g) as a lipid,Tween 80 (23.560 mg/g) as a surfactant, acetate (pH 5.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 84

Formulation 84 comprises phosphatidyl choline (54.000 mg/g) as a lipid,Tween 80 (36.000 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHA (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 85

Formulation 85 comprises phosphatidyl choline (50.000 mg/g) as a lipid,Tween 80 (40.000 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHA (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 86

Formulation 86 comprises phosphatidyl choline (48.611 mg/g) as a lipid,Tween 80 (38.889 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHA (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 87

Formulation 87 comprises phosphatidyl choline (46.575 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, acetate (pH 4) buffer. benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHA (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g). Example formulation 87 is an emulsion.

Example Formulation 88

Formulation 88 comprises phosphatidyl choline (46.575 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mglg) as an antimicrobial, BHA (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g). Example formulation 88 is suspension.

Example Formulation 89

Formulation 89 comprises phosphatidyl choline (46.575 mg/g) as a lipid,Tween 80 (3 8.425 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 90

Formulation 90 comprises phosphatidyl choline (50.000 mg/g) as a lipid,Tween 80 (40.000 mg/g) as a surfactant, acetate (pH 4.5) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mglg)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 91

Formulation 91 comprises phosphatidyl choline (94.444 mg/g) as a lipid,Tween 80 (75.556 mg/g) as a surfactant. acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 92

Formulation 92 comprises phosphatidyl choline (46.712 mg/g) as a lipid,Tween 80 (38.288 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 93

Formulation 93 comprises phosphatidyl choline (48.889 mg/g) as a lipid,Tween 80 (39.111 mg/g) as a surfactant, acetate (pH 4) buffer, benzylalcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200 mg/g)and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 94

Formulation 94 comprises phosphatidyl choline (39.721 mg/g) as a lipid,Tween 80 (50.279 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.25 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 95

Formulation 95 comprises phosphatidyl choline (90.000 mg/g) as a lipid,phosphate buffer (pH 6.5), benzyl alcohol or paraben as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 96

Formulation 96 comprises phosphatidyl choline (68.700 mg/g) as a lipid,Tween 80 (8.500 mg/g) as a surfactant, phosphate (pH 7.5) buffer, BHT(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, glycerol(30.000 mg/g), EDTA (1.000 mg/g) as a chelating agent, and ethanol(36.51 mg/g).

Example Formulation 97

Formulation 97 comprises phosphatidyl choline (71.460 mg/g) as a lipid,Tween 80 (4.720 mg/g) as a surfactant, phosphate (pH 7.5) buffer, BHA(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, glycerol(50.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(35.000 mg/g).

Example Formulation 98

Formulation 98 comprises phosphatidyl choline (71.460 mg/g) as a lipid,Tween 80 (4.720 mg/g) as a surfactant, phosphate (pH 7.8) buffer, BHA(0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, glycerol(15.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(35.000 mg/g).

Example Formulation 99

Formulation 99 comprises phosphatidyl choline (71.460 mg/g) as a lipid,Tween 80 (4.720 mg/g) as a surfactant, phosphate (pH 7.8) buffer, BHA(0.200 mg/g) and sodium metabi sulfite (0.500 mg/g) as antioxidants,glycerol (50.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (15.000 mg/g).

Example Formulation 100

Formulation 100 comprises phosphatidyl choline (71.4600 mg/g) as alipid, Tween 80 (4.720 mg/g) as a surfactant, phosphate (pH 7.5) buffer,BHA (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants,glycerol (50.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, andethanol (35.000 mg/g).

Example Formulation 101

Formulation 101 comprises phosphatidyl choline (46.575 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, phosphate (p11 4) buffer, BHT(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, andEDTA (3.000 mg/g) as a chelating agent. Example formulation 101 is anemulsion.

Example Formulation 102

Formulation 102 comprises phosphatidyl choline (46.575 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, phosphate (pH 4) buffer, BHT(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, andEDTA (3.000 mg/g). Example formulation 102 is a suspension.

Example Formulation 103

Formulation 103 comprises phosphatidyl choline (54.643 mg/g) as a lipid,Tween 80 (30.357 mg/g) as a surfactant, phosphate (pH 4) buffer, BHA(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, andEDTA (3.000 mg/g) as a chelating agent.

Example Formulation 104

Formulation 104 comprises phosphatidyl choline (39.72 mg/g)as a lipid,Tween 80 (50.279 mg/g) as surfactant, phosphate (pH 6.5) buffer, benzylalcohol or paraben (5.00 mg/g) as an antimicrobial, BHT (0.200 mg/g) andsodium metabisulfite (0.500 mg/g) as antioxidants, glycerol (30.000mg/g) as emollient, EDTA (3.000 mg/g) as the chelating agent, andethanol (30.000 mg/g).

Example Formulation 105

Formulation 105 comprises phosphatidyl choline (90.00 mg/g) as a lipid,phosphate (pH 6.5) buffer, benzyl alcohol or paraben as antimicrobial(5.000 mg/s), BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g) as emollient, EDTA (3.000 mg/g) asthe chelating agent, and ethanol (30.000 mg/g).

Example Formulation 106

Formulation 106 comprises phosphatidyl choline (46.57 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, phosphate (pH 4) buffer, BHT(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, andEDTA (3.000 mg/g) as the chelating agent. Formulation 106 is formulatedas an emulsion.

Example Formulation 107

Formulation 107 comprises phosphatidyl choline (46.57 mg/g) as a lipid,Tween 80 (38.425 mg/g) as a surfactant, phosphate (pH 4) buffer, BHT(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, andEDTA (3.000 mg/g) as the chelating agent. Formulation 107 as asuspension.

Example Formulation 108

Formulation 108 comprises phosphatidyl choline (54.64 mg/g)as a lipid,Tween 80 (30.357 mg/g) as a surfactant, phosphate (pH 4) buffer, BHA(0.500 mg/g) and sodium metabisulfite (0.200mg/g) as antioxidants, EDTA(3.000 mg/g) as the chelating agent.

Example Formulation 109

Formulation 109 comprises phosphatidyl glycerol and lysophospholipid(46.364 mg/g) as a lipid, Brij 98 (38.636 mg/g) as a surfactant, acetate(pH 4) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant. glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (25.000mg/g).

Example Formulation 110

Formulation 110 comprises phosphatidyl glycerol and lysophospholipid(46.364 mg/g) as a lipid, Brij 98 (38.636 mg/g) as a surfactant, acetate(pH 4) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, EDTA (3.000 mg/g) asa chelating agent, and ethanol (20.000 mg/g).

Example Formulation 111

Formulation 111 comprises phosphatidyl glycerol and lysophospholipid(46.098 mg/g) as a lipid, Brij 98 (43.902 mg/g) as a surfactant, acetate(pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, glycerol (15.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 112

Formulation 112 comprises phosphatidyl glycerol and lysophospholipid(43.537 mg/g) as a lipid, Brij 98 (41.463 mg/g) as a surfactant, acetate(pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, glycerol (30.000mg/g), and EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 113

Formulation 113 comprises phosphatidyl glycerol and lysophospholipid(45.000 mg/g) as a lipid, Brij 98 (45.000 mg/g) as a surfactant, acetate(pH 5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, EDTA (3.000 mg/g) asa chelating agent, and ethanol (30.000 mg/g).

Example Formulation 114

Formulation 114 comprises phosphatidyl glycerol and lysophospholipid(59.492 mg/g) as a lipid, Brij 98 (30.508 mg/g) as a surfactant, acetate(pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, glycerol (30.000mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol (30.000mg/g).

Example Formulation 115

Formulation 115 comprises phosphatidyl glycerol and lysophospholipid(39.054 mg/g) as a lipid, Brij 98 (45.946 mg/g) as a surfactant, acetate(pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, and EDTA (3.000 mg/g)as a chelating agent.

Example Formulation 116

Formulation 116 comprises phosphatidyl glycerol and lysophospholipid(35.854 mg/g) as a lipid, Brij 98 (34.146 mg/g) as a surfactant, acetate(pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) as an antioxidant, glycerol (30.000mg/g), and EDTA (3.000 mg/g) as a chelating agent.

Example Formulation 117

Formulation 117 comprises phosphatidyl choline and lysophospholipid(50.000 mg/g) as a lipid, Tween 80 (40.000 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 118

Formulation 118 comprises phosphatidyl choline and lysophospholipid(38.571 mg/g) as a lipid, Tween 80 (51.429 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g), and ethanol(30.000 mg/g).

Example Formulation 119

Formulation 119 comprises phosphatidyl choline and lysophospholipid(41.954 mg/g) as phospholipid, Tween 80 (50.546 mg/g) as surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g), and ethanol(30.000 mg/g).

Example Formulation 120

Formulation 120 comprises phosphatidyl choline and lysophospholipid(42.632 mg/g) as a lipid, Tween 80 (47.368 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 121

Formulation 121 comprises phosphatidyl choline and lysophospholipid(46.098 mg/g) as a lipid, Tween 80 (43.902 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 122

Formulation 122 comprises phosphatidyl choline and lysophospholipid(39.721 mg/g) as a lipid, Tween 80 (50.279 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 123

Formulation 123 comprises phosphatidyl choline and lysophospholipid(44.198 mg/g) as a lipid, Tween 80 (50.802 mg/g) as a surfactant,phosphate (pH 6.5) buffer. benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 124

Formulation 124 comprises phosphatidyl choline and lysophospholipid(46.453 mg/g) as a lipid, Tween 80 (51.047 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 125

Formulation 125 comprises phosphatidyl choline and lysophospholipid(51.221 mg/g) as a lipid. Tween 80 (43.779 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

Example Formulation 126

Formulation 126 comprises phosphatidyl choline (54.167 mg/g) as a lipid,Tween 80 (43.333 mg/g) as a surfactant, phosphate (pH 6.5) buffer,benzyl alcohol or paraben (5.000 mg/g) as an antimicrobial, BHT (0.200mg/g) and sodium metabisulfite (0.500 mg/g) as antioxidants, glycerol(30.000 mg/g), EDTA (3.000 mg/g) as a chelating agent, and ethanol(30.000 mg/g).

Example Formulation 127

Formulation 127 comprises phosphatidyl choline and lysophospholipid(66.440 mg/g) as a lipid, Brij 98 (23.560 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (3 0.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g). Example formulation 69 is an emulsion.

Example Formulation 128

Formulation 128 comprises phosphatidyl choline and lysophospholipid(66.440 mg/g) as a lipid, Brij 98 (23.560 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0.200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g). Example formulation 70 is asuspension.

Example Formulation 129

Formulation 129 comprises phosphatidyl choline and lysophospholipid(66.440 mg/g) as a lipid, Brij 98 (23.560 mg/g) as a surfactant,phosphate (pH 6.5) buffer, benzyl alcohol or paraben (5.000 mg/g) as anantimicrobial, BHT (0200 mg/g) and sodium metabisulfite (0.500 mg/g) asantioxidants, glycerol (30.000 mg/g), EDTA (3.000 mg/g) as a chelatingagent, and ethanol (30.000 mg/g).

It will be understood that the exact amounts of the components of theformula may be adjusted slightly without departing from the scope of theinvention. For example, in each of the above formulations, the amountantimicrobial be anywhere from about 1 mg/g to about 15 mg/g, or about 5m/g to about 12 mg/g, or 5.25 mg/g, 6, mg/6, 7 mg/g, 8 mg/g, 9 mg/g, 10mg/g, or 10.25 mg/g. Furthermore, the antimicrobial can be a combinationof ingredients, for example benzyl alcohol and parabenes (e.g., ethyland/or propyl).

Example Formulations 1 through 129 may also optionally includethickeners such as pectin. xanthan gum, HPMC gel, methylcellulose orcarbopol.

Example 2 DIRACTIN® Clinical Studies

Clinical studies directed to DIRACTIN® (ketoprofen in TRANSFERSOME® gel)were performed in the United States and in Europe. Table 9 providesdetails of the number of sites patients and the like in the European andUS studies.

TABLE 9 Comparative evaluation of various clinical studies conductedScreen PP Screen Ran- F. per Rel. Sites Failures dom PP Discont. Ratesite Discont. US 37 329 555 376/ 105 59% 10/12 19% (-06) 452 EU 71 1921399 1163 143 14% 16 10% (-03)

United States Study

A multicentre, randomized, double-blind, placebo-controlled study ofsafety and efficacy of epicutaneously applied DIRACTIN® (ketoprofen inTRANSFERSOME® gel) was conducted in the United states for the treatmentof osteoarthritis of the knee. The study was conducted from Q2/2008 toQ2/2009 and included 555 patients in 37 study centres. Patients weretreated twice daily at approximately 12 hour intervals for 12 weeks witheither:

-   -   (1) 100 mg ketoprofen (KT) in DIRACTIN® gel, epicutaneously        (e.c.) or    -   (2) Placebo gel with volume matched with 100 mg KT in DIRACTIN®,        e.c.

The DIRACTIN® gel and the placebo gel included transfersomes inaccordance with the descriptions provided herein. Rescue pain medicationwas provided in the amount of 500 mg acetaminophen up to four times aday, total 2 g.

The data from these studies established that the placebo transfersomes(i.e., transfersomes of the technology described herein) were active.Data from the US study are shown in FIGS. 1-5. FIG. 1 shows Mann-Whitneystatistics and continuous responder analysis of pain measurements inpatients. FIG. 2 provides the incidences of Adverse Events by organclass system. Subgroup analysis results for WOMAC pain intensity areshown in FIGS. 3 and 4; with FIG. 3 at a baseline of >5.5 and Figure forat baseline≦5.5. Descriptive statistics for the use of rescue medicationare provided in FIG. 5.

European Study

A multicentre, randomized, double-blind, placebo-controlled study ofsafety and efficacy of epicutaneously applied DIRACTIN® (ketoprofen inTRANSFERSOME® gel) was also conducted in Europe for the treatment ofosteoarthritis of the knee. The study was conducted from Q2/2008 toQ2/2009 and included 1,399 patients in 71 study centres (in CzechRepublic, Germany, Poland, and UK). Patients were treated twice daily atapproximately 12 hour intervals for 12 weeks with either:

-   -   (1) 50 mg ketoprofen (KT) in DIRACTIN® gel, epicutaneously        (e.c.)    -   (2) Placebo gel with volume matched with 50 mg KT in DIRACTIN®,        e.c.    -   (3) 100 mg ketoprofen (KT) in DIRACTIN® gel, e.c.    -   (4) Placebo gel with volume matched with 100 mg KT in DIRACTIN®,        e.c.    -   (5) 100 mg celecoxib capsule (CELEBREX®, Pfizer), oral    -   (6) Placebo capsule, matching celecoxib, oral

The DIRACTIN® gel and the placebo gel included transfersomes inaccordance with the descriptions provided herein. Rescue pain medicationwas provided in the amount of 500 mg acetaminophen up to four times aday, total 2 g.

A schematic illustrating the study design is shown on FIG. 6 and anillustration showing the statistical evaluations are shown in FIG. 7.

The data from these studies established that the placebo transfersomes(i.e., transfersomes of the technology described herein) were active.Data from the European study are shown in FIGS. 8-17. FIGS. 8-9 showMann-Whitney statistics and continuous responder analysis of painmeasurements in patients, with FIG. 8 showing results for the 100 mgdose of KT and placebo gel and FIG. 9 showing results for the 50 mg doseof KT and placebo gel, FIGS. 10 and 11 show WOMAC pain and Mann-Whitneystatistics for the 100 mg celecoxib capsule (CELEBREX®, Pfizer) and theoral placebo. FIG. 12 shows Mann-Whitney statistics and continuousresponder analysis of pain measurements in patients for the 50 mg doseof KT vs. the 100 mg celecoxib capsule (CELEBREX®, Pfizer). FIG. 13shows Mann-Whitney statistics for the 100 mg dose of KT vs. the oralplacebo and FIG. 14 shows Mann-Whitney statistics for the 50 mg dose ofKT vs. the oral placebo. FIG. 15 shows Mann-Whitney statistics for the100 mg dose of KT vs. 100 mg celecoxib capsule (CELEBREX®, Pfizer). FIG.16 shows Mann-Whitney statistics for the 100 mg dose of KT vs. the oralplacebo, using the “per protocol population” instead of “intention totreat” (ITT). FIG. 17 shows possibly treatment related adverse events(Aes) observed in the study.

Inter-Studies Results and Conclusions

The combined results of the US and European studies establish that theplacebo gel (transfersomes gel) is equivalent or better than theketoprofen-containing gel, and that each have similar efficacy to oralcelecoxib. All three of these products are shown to be superior to oralplacebo. The effectiveness of the placebo gel (transfersomes gel) wassimilar to oral drugs acting on Cox-II but had substantially lower sideeffects liability. FIG. 18 shows the average group change for thevarious treatments at week 6 and 12 of the studies. FIG. 19 shows theeffect of the placebo response in published data and by the placebo gelthat is the subject of the invention.

Example 3 Clinical Trials for Transfersomes for Inflammatory Dermatoses

A multicentre, randomized, double-blind, placebo-controlled study ofsafety and efficacy of IDEA-070 (transfersome spray containingketoprofen) was conducted in the Germany for the treatment ofinflammatory dermatological diseases. The study was conducted fromQ1/2005 to Q4/2005 and included 240 patients in 7 study centres. Thetarget indications for the study were atopic eczema, dishydrotic handeczema, plaque type psoriasis, seborrheic eczema, and acne vulgaris. Thetreatment group used in the study received 0.24 mg ketoprofen per cm²skin in IDEA-070 spray, e.c., b.i.d. and the controls received a topicalplacebo (randomisation: IDEA-070/placebo: 2/1). Transfersomes used inthis study were made in accordance with transfersomes as describedherein. The primary objectives of the study were to evaluate the effectsof IDEA-070 compared to a placebo in patients with atopic eczema,dishydrotic hand eczema, plaque type psoriasis, seborrheic eczema, andacne vulgaris using the Investigator Global Assessments score (IGA) andthe secondary objectives were to evaluate the efficacy of IDEA-070compared to a placebo using the Patient Global Assessments score (PGA)and indication specific scores (SCORAD, DASI, PASI, GAGS) as well as totest the safety of IDEA-070.

Patient inclusion in the study required that the patient (1) had one ofthe following diseases (mild to moderate): atopic eczema, dishydrotichand eczema, plaque type psoriasis (hyperkeratoses removed beforetreatment by urea or salicylic acid), seborrheic eczema in the face orhead), or acne vulgaris; (2) was aged 18-80 years; and (3) that women ofchildbearing potential were using a reliable method of contraception.Patients meeting any of the following criteria were excluded from thestudy: Systemic therapy for skin diseases within 2 weeks prior to startof treatment; UV therapy within 4 weeks prior to start of treatment;Chronic or acute illness requiring systemic anti-inflammatory treatment;Skin cancer and precancerous skin lesions; History of peptic ulcers orgastric intolerance with NSAIDs; History of asthma bronchiale; Historyof chronic airway infection; History of renal insufficiency;Thrombocytopathia; Immunosuppressants (e.g., corticosteroids) within 2weeks prior to start of treatment; Known sensitisation to NSAIDs;Pregnancy or lactation; Mental disorders.

Table 10 summarizes the study population used in the study.

TABLE 10 Placebo IDEA-070 All Acne 17 33 50 Atopic E. 17 31 48Dishydrotic E. 18 34 52 Psoriasis 16 35 51 Seborrheic E. 14 34 48 Total249

The data from these studies established that the placebo transfersomes(i.e., transfersomes of the technology described herein) were active.FIGS. 20-23 provide results of the study. FIG. 20 shows the IGA scorefor the five main objective indications. As shown of FIG. 20 there wasno significant difference between IDEA-070 and placebo for any of theindications except for seborrheic eczema, in which the placebo gelshowed a more marked improvement that TDT 070 with a p value of p=0.031.FIGS. 21-23 show the IGA scores at week 1, 2 and 3 for seborrheiceczema, psoriasis, psoriasis area and severity index (PASI), anderythema, respectively.

Adverse events by indication observed in the study are shown in Table11.

TABLE 11 Placebo IDEA-070 Akne 17.6% 24.2% Atopic E. 41.2% 32.3%Dishydrotic E. 16.7% 20.6% Psoriasis 12.5% 25.7% Seborrheic E. 7.1%23.5% All 19.5% 25.1%

Adverse events by system organ class-skin and subcutaneous tissue areshown in Table 12.

TABLE 12 Placebo IDEA-070 Akne 0 1 Atopic E. 0 5 Dishydrotic E. 0 1Psoriasis 1 4 Seborrheic E. 0 0 All 1 11

Example 4 Sequestration of Arachidonic Acid

Assays were performed to assess the effect of transfersomes arachidonicacid sequestration. The assays used in these studies involved usingarachidonic acid as a substrate and measuring cylclooxygenase activityas an indicator of arachidonic acid sequestration Other assays could beused to measure/analyze arachidonic acid sequestration. For examplesequestration could be measured using radiolabeled arachidonic acid (oranother lipid), adding the transfersomes to sequester the lipid,centrifuging to separate the transfersomes (or using another method ofseparating transfersomes, such as filtration) and measuring theradiolabeled lipid in the transfersomes as an indicator ofsequestration.

Transfersomes used in this study were made in accordance with thetransfersomes described herein.

Representative results from the COX inhibition/Sequestration assays areshown in FIGS. 25-26. FIG. 25 shows a double reciprocal plot of COXreaction velocity versus arachidonic acid concentration for a control,50 μM ketoprofen and 100 μM transfersomes. The differences between theresults of the control and the 100 μM transfersomes indicatesequestration of the arachidonic acid substrate. FIG. 26 shows the COXreaction velocity using 16 μM arachidonic acid and increasing amounts oftransfersomes. The saturation at between 50-70 μm transfersomes suggeststhat the critical sequestration capacity of the transfersomes occurs inthis range (as marked on the figure).

Example 5 Sequestration of Other Lipids

Assays are performed to assess the effect of transfersomes on otherlipids. Other lipids include cholesterol, mediators of pain andinflammation (e.g., such as prostaglandins, prostaglandin precursors andleukotrienes. The transfersomes are effective in sequestering identifiedlipids.

1-122. (canceled)
 123. A vesicular formulation for use in the treatmentof inflammatory dermatoses disorders, gout or fungal infectioncomprising one or more phospholipids and one or more surfactants, in theabsence of a pharmaceutically active agent.
 124. A vesicular formulationas claimed in claim 1, for use in the treatment of atopic eczema,dishydrotic hand eczema, plaque type psoriasis, seborrheic eczema, oracne vulgaris.
 125. A vesicular formulation of claim 1, wherein theformulation is a cream, lotion, ointment, gel, solution, spray, lacqueror film forming solution.
 126. A vesicular formulation as claimed inclaim 1, wherein the ratio of phospholipid to surfactant is about 1:3 toabout 30:1.
 127. A vesicular formulation, as claimed in claim 1, whereinthe formulation contains 2.0-10.0% by weight phospholipid.
 128. Avesicular formulation, as claimed in claim 1, wherein the formulationcontains 1.0-5.0% by weight surfactant.
 129. A vesicular formulation, asclaimed in claim 1, wherein the formulation contains 0.2% to about 0.5%by weight surfactant.
 130. A vesicular formulation, as claimed in claim1, wherein the phospholipid is phosphatidylcholine.
 131. A vesicularformulation, as claimed in claim 1, wherein the surfactant is a nonionicsurfactant selected from the group consisting of: polyoxyethylenesorbitans, polyhydroxyethylene stearates or polyhydroxethylenelaurylethers.
 132. A vesicular formulation, as claimed in claim 1,wherein the surfactant is polysorbate 80 (Tween 80).